Home About us Contact
|
Home About us Contact | ||
|
|
||
Myoglobin
Kinds of Myoglobin Terms modified by Myoglobin Selected AbstractsExtending the Pressure,Temperature State Diagram of MyoglobinHELVETICA CHIMICA ACTA, Issue 3 2005Filip Meersman The pressure,temperature (P,T) diagram of proteins proposed by Hawley concerns the equilibrium between native and denatured forms. However, the importance of protein aggregation is increasingly recognized, and it has been suggested that certain aggregated states represent alternative folds of the polypeptide chain. Here, we present a P,T -diagram for myoglobin in which we include the aggregated state and suggest to call it a P,T -state diagram, as not all boundaries are true equilibrium transitions. We observe by Fourier transform infrared spectroscopy that increasing temperature causes the protein to aggregate, but that a subsequent further temperature increase results in the dissociation of this aggregate. Moreover, we observe that moderate pressures stabilize myoglobin against thermal denaturation. We hypothesize that this effect originates from the volume changes associated with the aggregation transition. [source] Evaluation of stratus® CS Stat fluorimetric analyser for measurement of cardiac markers Troponin I (cTnI), creatine kinase MB (CK-MB), and myoglobinJOURNAL OF CLINICAL LABORATORY ANALYSIS, Issue 6 2001Bénédicte Bénéteau-Burnat Abstract Myoglobin, CK-MB, and Troponin I (cTnI) are cardiac muscle necrosis markers that are useful for detecting acute myocardial infarction (AMI). The Stratus® CS (Dade Behring, Inc.) is a discrete fluorimetric immunoassay analyser designed for the determination of the three cardiac markers from a single sample of whole blood or plasma. Overall analytical performances of the Stratus® CS provided by Dade Behring were evaluated according to the French Society of Clinical Biology guidelines. Within-run imprecision (n = 20) for the three parameters at three levels gave values under 5%, whereas CVs for between-run imprecision (n = 20) were under 6%. The sensitivities were 0.03 ,g/L for cTnI and 0.4 ,g/L for CK-MB. Linearities extended from 0,50 ,g/L for cTnI, 0,140 ,g/L for CK-MB, and 1,900 ,g/L for myoglobin. The results, particularly those obtained on whole-blood samples, correlated well with those obtained on Stratus® II. We did not find any interference with haemolysis, icterus, or lipemia. The system was very easy to use, and fulfills the requirements for the analysis of the three cardiac markers in patients with acute chest pain in emergency situations. J. Clin. Lab. Anal. 15:314,318, 2001. © 2001 Wiley-Liss, Inc. [source] The Complex of Apomyoglobin with the Fluorescent Dye Coumarin 153,PHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 5 2004P. K. Chowdhury ABSTRACT Understanding a protein's dielectric response requires both a theoretical model and a well-defined experimental system. The former has already been proposed by Song (J. Chem. Phys. 116, 9359 [2002]). We suggest that the latter is provided by the complex of coumarin 153 (C153) with apomyoglobin (ApoMb). C153 has been exhaustively studied and has proven to be an excellent probe of the solvation dynamics of polar solvents. Myoglobin is one of the most thoroughly studied proteins. Myoglobins from a wide range of species have been subject to X-ray structural analysis and site-directed mutagenesis. Here, we demonstrate the existence of a robust C153-apomyglobin system by means of molecular dynamics simulations, equilibrium binding studies using a Job's plot and capillary electrophoresis, circular dichroism and time-resolved fluorescence. The reorganization energy of C153 bound to ApoMb is compared with that of C153 in bulk solvent using the method of Jordanides et al. (J. Phys. Chem. B 103, 7995 [1999]). [source] Optimization of Electrochemical and Peroxide-Driven Oxidation of Styrene with Ultrathin Polyion Films Containing Cytochrome P450cam and MyoglobinCHEMBIOCHEM, Issue 1 2003Bernard Munge Abstract The catalytic and electrochemical properties of myoglobin and cytochrome P450camin films constructed with alternate polyion layers were optimized with respect to film thickness, polyion type, and pH. Electrochemical and hydrogen peroxide driven epoxidation of styrene catalyzed by the proteins was used as the test reaction. Ionic synthetic organic polymers such as poly(styrene sulfonate), as opposed to SiO2nanoparticles or DNA, supported the best catalytic and electrochemical performance. Charge transport involving the iron heme proteins was achieved over 40,320 nm depending on the polyion material and is likely to involve electron hopping facilitated by extensive interlayer mixing. However, very thin films (ca. 12,25 nm) gave the largest turnover rates for the catalytic epoxidation of styrene, and thicker films were subject to reactant transport limitations. Classical bell-shaped activity/pH profiles and turnover rates similar to those obtained in solution suggest that films grown layer-by-layer are applicable to turnover rate studies of enzymes for organic oxidations. Major advantages include enhanced enzyme stability and the tiny amount of protein required. [source] Noncovalent Modulation of pH-Dependent Reactivity of a Mn,Salen Cofactor in Myoglobin with Hydrogen PeroxideCHEMISTRY - A EUROPEAN JOURNAL, Issue 30 2009Jun-Long Zhang Dr. Abstract To demonstrate protein modulation of metal-cofactor reactivity through noncovalent interactions, pH-dependent sulfoxidation and 2,2,-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) oxidation reactivity of a designed myoglobin (Mb) containing non-native Mn,salen complex (1) was investigated using H2O2 as the oxidant. Incorporation of 1 inside the Mb resulted in an increase in the turnover numbers through exclusion of water from the metal complex and prevention of Mn,salen dimer formation. Interestingly, the presence of protein in itself is not enough to confer the increase activity as mutation of the distal His64 in Mb to Phe to remove hydrogen-bonding interactions resulted in no increase in the turnover numbers, while mutation His64 to Arg, another residue with ability to hydrogen-bond interactions, resulted in an increase in reactivity. These results strongly suggest that the distal ligand His64, through its hydrogen-bonding interaction, plays important roles in enhancing and fine-tuning reactivity of the Mn,salen complex. Nonlinear least-squares fitting of rate versus pH plots demonstrates that 1,Mb(H64X) (X=H, R and F) and the control Mn,salen 1 exhibit pKa values varying from pH,6.4 to 8.3, and that the lower pKa of the distal ligand in 1,Mb(H64X), the higher the reactivity it achieves. Moreover, in addition to the pKa at high pH, 1,Mb displays another pKa at low pH, with pKa of 5.0±0.08. A comparison of the effect of different pH on sulfoxidation and ABTS oxidation indicates that, while the intermediate produced at low pH conditions could only perform sulfoxidation, the intermediate at high pH could oxidize both sulfoxides and ABTS. Such a fine-control of reactivity through hydrogen-bonding interactions by the distal ligand to bind, orient and activate H2O2 is very important for designing artificial enzymes with dramatic different and tunable reactivity from catalysts without protein scaffolds. [source] On the Influence of the Local Environment on the CO Stretching Frequencies in Native Myoglobin: Assignment of the B-States in MbCOCHEMPHYSCHEM, Issue 10 2006Markus Meuwly Prof. Dr. Frequency shifts: The influence of the local electrostatic environment on the ground vibrational level of photodissociated CO in native myoglobin is investigated by performing QM/MM calculations at the DFT level. The distribution of CO frequencies in the FeCO conformation occurs at higher wavenumbers than in the FeOC conformation (see figure). [source] The Complex of Apomyoglobin with the Fluorescent Dye Coumarin 153,PHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 5 2004P. K. Chowdhury ABSTRACT Understanding a protein's dielectric response requires both a theoretical model and a well-defined experimental system. The former has already been proposed by Song (J. Chem. Phys. 116, 9359 [2002]). We suggest that the latter is provided by the complex of coumarin 153 (C153) with apomyoglobin (ApoMb). C153 has been exhaustively studied and has proven to be an excellent probe of the solvation dynamics of polar solvents. Myoglobin is one of the most thoroughly studied proteins. Myoglobins from a wide range of species have been subject to X-ray structural analysis and site-directed mutagenesis. Here, we demonstrate the existence of a robust C153-apomyglobin system by means of molecular dynamics simulations, equilibrium binding studies using a Job's plot and capillary electrophoresis, circular dichroism and time-resolved fluorescence. The reorganization energy of C153 bound to ApoMb is compared with that of C153 in bulk solvent using the method of Jordanides et al. (J. Phys. Chem. B 103, 7995 [1999]). [source] The effect of hypoxia on pulmonary O2 uptake, leg blood flow and muscle deoxygenation during single-leg knee-extension exerciseEXPERIMENTAL PHYSIOLOGY, Issue 3 2004Darren S. DeLorey The effect of hypoxic breathing on pulmonary O2 uptake (VO2p), leg blood flow (LBF) and O2 delivery and deoxygenation of the vastus lateralis muscle was examined during constant-load single-leg knee-extension exercise. Seven subjects (24 ± 4 years; mean ±s.d.) performed two transitions from unloaded to moderate-intensity exercise (21 W) under normoxic and hypoxic (PETO2= 60 mmHg) conditions. Breath-by-breath VO2p and beat-by-beat femoral artery mean blood velocity (MBV) were measured by mass spectrometer and volume turbine and Doppler ultrasound (VingMed, CFM 750), respectively. Deoxy-(HHb), oxy-, and total haemoglobin/myoglobin were measured continuously by near-infrared spectroscopy (NIRS; Hamamatsu NIRO-300). VO2p data were filtered and averaged to 5 s bins at 20, 40, 60, 120, 180 and 300 s. MBV data were filtered and averaged to 2 s bins (1 contraction cycle). LBF was calculated for each contraction cycle and averaged to 5 s bins at 20, 40, 60, 120, 180 and 300 s. VO2p was significantly lower in hypoxia throughout the period of 20, 40, 60 and 120 s of the exercise on-transient. LBF (l min,1) was approximately 35% higher (P > 0.05) in hypoxia during the on-transient and steady-state of KE exercise, resulting in a similar leg O2 delivery in hypoxia and normoxia. Local muscle deoxygenation (HHb) was similar in hypoxia and normoxia. These results suggest that factors other than O2 delivery, possibly the diffusion of O2, were responsible for the lower O2 uptake during the exercise on-transient in hypoxia. [source] Effects of Ischaemia on Subsequent Exercise-Induced Oxygen Uptake Kinetics in Healthy Adult HumansEXPERIMENTAL PHYSIOLOGY, Issue 2 2002Michael L. Walsh Leg muscles were occluded (33 kPa) prior to exercise to determine whether the induced metabolic changes, and reactive hyperaemia upon occlusion release just prior to the exercise, would accelerate the subsequent oxygen consumption (V,O2) response. Eight subjects performed double bouts (6 min duration, 6 min rest in-between) of square wave leg cycle ergometry both below and above their lactate threshold (LT). Prior to exercise, large blood pressure cuffs were put around the upper thighs. Occlusion durations were 0 min (control), 5 min and 10 min. Ischaemia was terminated within 5 s prior to exercise onset. Heart rate, V,O2, ventilatory rate (V,E), electromyogram (EMG) and haemoglobin/myoglobin (Hb/Mb) saturation were recorded continuously. Single exponential modelling demonstrated that, compared to control (time constant = 53.9 ± 13.9 s), ischaemia quickened the V,O2 response (P < 0.05) for the first bout of exercise above LT (time constant = 48.3 ± 14.5 s) but not to any other exercise bout below or above LT. The 3-6 min integrated EMG (iEMG) slope was correlated to the 3-6 min V,O2 slope (r = 0.73). Hb/Mb saturation verified the ischaemia but did not show a consistent relation to the V,O2 time course. Reactive hyperaemia induced a faster V,O2 response for work rates above LT. The effect, while significant, was not large considering the expected favourable metabolic and circulatory changes induced by ischaemia. [source] Direct Electrochemistry and Electrocatalysis of Myoglobin Immobilized on Gold Nanoparticles/Carbon Nanotubes Nanohybrid FilmELECTROANALYSIS, Issue 17 2008Wei Cao Abstract A novel nanohybrid material, constructed by gold nanoparticles (GNPs) and multiwalled carbon nanotubes (MWNTs), was designed for immobilization and biosensing of myoglobin (Mb). Morphology of the nanohybrid film was characterized by SEM. UV-vis spectroscopy demonstrated that Mb on the composite film could retain its native structure. Direct electrochemistry of Mb immobilized on the GNPs/MWNTs film was investigated. The immobilized Mb showed a couple of quasireversible and well-defined cyclic voltammetry peaks with a formal potential of about ,0.35,V (vs. Ag/AgCl) in pH,6.0 phosphate buffer solution (PBS) solution. Furthermore, the modified electrode also displayed good sensitivity, wide linear range and long-term stability to the detection of hydrogen peroxide. The experiment results demonstrated that the hybrid matrix provided a biocompatible microenvironment for protein and supplied a necessary pathway for its direct electron transfer. [source] Microchip isoelectric focusing with monolithic immobilized pH gradient materials for proteins separationELECTROPHORESIS, Issue 23 2009Yu Liang Abstract Monolithic immobilized pH gradient (M-IPG) materials were prepared in microchannles by photoinitiated polymerization of acrylamide, glycidylmethacrylate and Bis, followed by the attachment of focused Ampholine onto the surface of porous monoliths via epoxide groups. With M-IPG materials as matrix, FITC-labeled ribonuclease B, myoglobin and ,-casein were well separated by microchip isoelectric focusing (,CIEF) without carrier amphocytes (CAs) added in the buffer. Both chemical and pressure mobilization were applied to drive focused zones for LIF detection. Our experimental results showed that pressure mobilization was preferable with neglectable band broadening, and good peak shape and high detection sensitivity were obtained. All these results demonstrate that ,CIEF with M-IPG materials is not only an efficient mode for protein enrichment and separation but also attractive to couple with other CE modes to achieve multi-dimensional separation or MS for further identification, without the interference of mobile CAs. [source] Novel negatively charged tentacle-type polymer coating for on-line preconcentration of proteins in CEELECTROPHORESIS, Issue 4 2009Liang Xu Abstract A novel negatively charged tentacle-type polymer-coated capillary column was fabricated and applied for on-line extraction and preconcentration of proteins. The polymer coating was prepared by glycidyl-methacrylate graft polymerization in a silanized capillary column and the following sulfonic acid group functionalization. It had high surface area and offered high phase ratio for protein adsorption. In addition, the polymer-coated capillary column provided more stable EOF than a bare uncoated capillary. These features of the polymer coating facilitated the extraction of proteins through electrostatic interactions. This was used to extract proteins. The extracted analytes were then desorbed and focused by EOF in the direction opposite to the sample injection flow for subsequent CE. With this procedure, over 1500-fold sensitivity enhancement was realized for myoglobin (MB) as compared with a normal capillary zone electrophoresis. By comparison of the peak areas of the enriched protein, it was found that the polymer-coated column could capture proteins about 30 times more than the uncoated column. In addition, the separation of a protein mixture containing 0.4,,g/mL of MB and 0.4,,g/mL of insulin was demonstrated by the on-line preconcentration and electrophoretic separation with the polymer-coated column. [source] High-efficiency protein extraction from polyacrylamide gels for molecular mass measurement by matrix-assisted laser desorption/ionization-time of flight-mass spectrometryELECTROPHORESIS, Issue 6 2005Ya Jin Abstract A simple and fast method of protein extraction from Coomassie Brilliant Blue (CBB)-stained polyacrylamide gels suited for molecular mass measurement of proteins by matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS) is reported. Proteins in CBB-stained gel pieces were extracted by a 10-min soaking in 0.1,M NaOH at 25°C. The recovery of this one-step extraction method was 34,73% for proteins <67,kDa. CBB adduction to proteins during mass spectrometric analysis was avoided by a destaining step before the alkaline extraction. The molecular mass values of the extracted proteins coincided with those of purified proteins within ±0.01,0.10% deviation for all the proteins <36,kDa. Because of the high extraction recovery, mass measurement was possible for the proteins extracted from CBB-stained gels with loaded protein quantities as little as 34,ng for cytochrome,c, ,-lactalbumin, myoglobin, ,-lactoglobulin, trypsinogen, and carbonic anhydrase (12.4,29.0,kDa), 340,ng for glyceraldehyde-3-phosphate dehydrogenase (35.6,kDa) and albumin (66.3,kDa). This method provides a highly efficient approach to utilize CBB-stained one- or two-dimensional gels for whole protein analysis using MALDI-TOF-MS. [source] Application of dodecyldimethyl (2-hydroxy-3-sulfopropyl) ammonium in wall modification for capillary electrophoresis separation of proteinsELECTROPHORESIS, Issue 3 2005Wei Wei Abstract A zwitterionic surfactant, dodecyldimethyl (2-hydroxy-3-sulfopropyl) ammonium (C12H25N+(CH3)2CH2CHOHCH2SO3,), named dodecyl sulfobetaine (DSB), was used as a novel modifier to coat dynamically capillary walls for capillary electrophoresis separation of basic proteins. The DSB coating suppressed the electroosmotic flow (EOF) in the pH range of 3,12. At high DSB concentration, the EOF was suppressed by more than 8.8,times. The DSB coating also prevented successfully the adsorption of cationic proteins on the capillary wall. Anions, such as Cl,, Br,, I,, SO42,, CO32,, and ClO4,, could be used as running buffer modifiers to adjust the EOF for better separation of analytes. Using this dynamically coated capillary, a mixture of eight inorganic anions achieved complete separation within 4.2,min with the efficiencies from 24,000 to 1,310,000,plates/m. In the presence of ClO4, as EOF adjustor, the separation of a mixture containing four basic proteins (lysozyme, cytochrome c, ,-chymotrypsinogen,A, and myoglobin) yielded efficiencies of 204,000,896,000,plates/m and recoveries of 88%,98%. Migration time reproducibility of these proteins was less than 0.5% relative standard deviation (RSD) from run to run and less than 3.1% RSD from day to day, showing promising application of this novel modifier in protein separation. [source] Quantification of myoglobin deoxygenation and intracellular partial pressure of O2 during muscle contraction during haemoglobin-free medium perfusionEXPERIMENTAL PHYSIOLOGY, Issue 5 2010Hisashi Takakura Although the O2 gradient regulates O2 flux from the capillary into the myocyte to meet the energy demands of contracting muscle, intracellular O2 dynamics during muscle contraction remain unclear. Our hindlimb perfusion model allows the determination of intracellular myoglobin (Mb) saturation () and intracellular oxygen tension of myoglobin () in contracting muscle using near infrared spectroscopy (NIRS). The hindlimb of male Wistar rats was perfused from the abdominal aorta with a well-oxygenated haemoglobin-free Krebs,Henseleit buffer. The deoxygenated Mb (,[deoxy-Mb]) signal was monitored by NIRS. Based on the value of ,[deoxy-Mb],,,and,,were calculated, and the time course was evaluated by an exponential function model. Both,,and,,started to decrease immediately after the onset of contraction. The steady-state values of,,and,,progressively decreased with relative work intensity or muscle oxygen consumption. At the maximal twitch rate,,,and,,were 49% and 2.4 mmHg, respectively. Moreover, the rate of release of O2 from Mb at the onset of contraction increased with muscle oxygen consumption. These results suggest that at the onset of muscle contraction, Mb supplies O2 during the steep decline in,, which expands the O2 gradient to increase the O2 flux to meet the increased energy demands. [source] Proximal ligand motions in H93G myoglobinFEBS JOURNAL, Issue 19 2002Stefan Franzen Resonance Raman spectroscopy has been used to observe changes in the iron,ligand stretching frequency in photoproduct spectra of the proximal cavity mutant of myoglobin H93G. The measurements compare the deoxy ferrous state of the heme iron in H93G(L), where L is an exogenous imidazole ligand bound in the proximal cavity, to the photolyzed intermediate of H93G(L)*CO at 8 ns. There are significant differences in the frequencies of the iron,ligand axial out-of-plane mode ,(Fe,L) in the photoproduct spectra depending on the nature of L for a series of methyl-substituted imidazoles. Further comparison was made with the proximal cavity mutant of myoglobin in the absence of exogenous ligand (H93G) and the photoproduct of the carbonmonoxy adduct of H93G (H93G-*CO). For this case, it has been shown that H2O is the axial (fifth) ligand to the heme iron in the deoxy form of H93G. The photoproduct of H93G-*CO is consistent with a transiently bound ligand proposed to be a histidine. The data presented here further substantiate the conclusion that a conformationally driven ligand switch exists in photolyzed H93G-*CO. The results suggest that ligand conformational changes in response to dynamic motions of the globin on the nanosecond and longer time scales are a general feature of the H93G proximal cavity mutant. [source] Effects of pressure on the activity and spectroscopic properties of carboxyl proteinasesFEBS JOURNAL, Issue 3 2001Apparent correlation of pepstatin-insensitivity, pressure response The pressure dependence of the activity and spectroscopic properties of four carboxyl proteinases were investigated. Two were pepstatin-sensitive carboxyl proteinases (porcine pepsin and proteinase A from baker's yeast) and two were pepstatin-insensitive carboxyl proteinases (from Pseudomonas sp. 101 (pseudomonapepsin; PCP) and Xanthomonas sp. T-22 (xanthomonapepsin; XCP)). The specificity constant [kcat/Km(app)] of PCP and XCP for a synthetic peptide substrate showed only a slight decrease with increasing pressure, whereas pepsin and proteinase A showed substantial disactivation at higher pressures. The calculated apparent activation volume (,Vkcat/Km) was about 1, 3, 13, and 14 mL·mol,1 for PCP, XCP, pepsin, and proteinase A, respectively. The hydrolysis of acid-denatured myoglobin by the four carboxyl proteinases was only slightly affected by high pressure (except for proteinase A at 400 MPa), in contrast to the results for the peptide hydrolysis. In fact, PCP, XCP, and proteinase A actually showed slightly higher degradations of acid-denatured myoglobin at higher pressures. The residual activities of these enzymes after the incubation at high pressures implied a pressure-induced stabilization towards autolysis. The changes in the fourth derivative near-UV absorbance spectrum of the four enzymes in aqueous solution were measured at various pressures from 0.1 to 400 MPa. Upon an increase in pressure, the peaks from PCP and XCP red-shifted slightly, whereas pepsin and proteinase A blue-shifted substantially, thus indicating a more polar environment. The intrinsic fluorescence also decreased upon increasing pressure. However, the change for XCP was rather small, but the change for the other three was very large. The changes in the peak wavelength for pepsin and proteinase A were characteristic, and also indicated a more polar environment under high pressure. An analysis by the center of spectra mass (CSM) gave the ,G and ,V of transition as 9.8 kJ·mol,1 and ,24 mL·mol,1 (pepsin) and 11.7 kJ·mol,1 and ,43 mL·mol,1 (proteinase A), respectively, by assuming a simple two-state transition. The circular dichroism (CD) showed relatively small changes after 1-h incubations at 400 MPa, indicating that the secondary structures were largely maintained. [source] Cardioprotection of cariporide evaluated by plasma myoglobin and troponin I in myocardial infarction in pigsFUNDAMENTAL & CLINICAL PHARMACOLOGY, Issue 2 2006Robert Létienne Abstract The cardioprotective effects of cariporide were investigated against myoglobin and troponin I elevation in a model of myocardial infarction in pig, and the possible relationship between these markers and myocardial infarct size. The left circumflex coronary artery was ligated for 60-min and then reperfused for 48-h. Plasma levels of myoglobin and troponin I were quantified during reperfusion. Vehicle or cariporide (2.5 mg/kg) were administered i.v. before ischaemia and infused throughout ischaemia and for the beginning of reperfusion. In vehicle-treated pigs, the infarct size represented 26% ± 3% of the area at risk. Cariporide significantly decreased the infarct size by 66% ± 9%, and significantly reduced plasma levels of myoglobin and troponin I. A strongly correlated linear relationship between myocardial necrosis and plasma levels of myoglobin (R = 0.966, P < 0.0001) or troponin I (R = 0.855, P < 0.0001) was clearly identified. In conclusion, in our porcine model of myocardial infarction, even with small infarcts (in the presence of cariporide), plasma levels of myoglobin and troponin I are predictive of the presence of necrosis and its extent. [source] Polymer Films Composed of Surface-Bound Nanofilaments with a High Aspect Ratio, Molecularly Imprinted with Small Molecules and ProteinsADVANCED FUNCTIONAL MATERIALS, Issue 8 2009Ana Valvanuz Linares Abstract Hierarchically nanostructured materials that combine two or more levels of structuring and that exhibit a combination of useful features have gained considerable interest over recent years. Here, the generation of surface-bound nanofilaments with a high aspect ratio by nanomolding on a nanoporous template surface is described. The filaments, at the same time, carry molecularly imprinted binding sites. The dye fluorescein and the protein myoglobin are used as model templates for imprinting. The surfaces exhibit specific binding as revealed by fluorescence microscopy. The wetting properties of the surfaces depend on the dimensions of the nanofilaments and on the nature of the polymer. It is believed that these materials can potentially be useful for applications in biosensors and biochips. [source] Adult extracardiac rhabdomyoma: Light and immunohistochemical studies of two cases in the parapharyngeal spaceHEAD & NECK: JOURNAL FOR THE SCIENCES & SPECIALTIES OF THE HEAD AND NECK, Issue 3 2006Kristine Bjřrndal Sřrensen MD Abstract Background. We present two cases of adult rhabdomyoma in the parapharyngeal space. They are rare benign tumors with a characteristic histologic appearance. Methods. The tumors were studied by light and immunohistochemical analysis using stains characteristic of striated muscle fibers. Results. Cross-striation was demonstrated by phosphotungstic acid hematoxylin (PTAH), muscle specific actin, desmin, and myoglobin while dystrophin was expressed in the cell membranes. Clonal origin was confirmed by expression of myosin heavy chain-fast only. Expression of myosin-neonatal and myogenin proved slight proliferation with incipient differentiation in an otherwise mature tumor. Conclusion. The head and neck area harbors 90% of adult rhabdomyomas and should be considered in a differential diagnosis in this region. Immunohistochemistry confirms that the tumors are almost totally mature neoplasms of clonal origin. © 2006 Wiley Periodicals, Inc. Head Neck28: XXX,XXX, 2006 [source] Extending the Pressure,Temperature State Diagram of MyoglobinHELVETICA CHIMICA ACTA, Issue 3 2005Filip Meersman The pressure,temperature (P,T) diagram of proteins proposed by Hawley concerns the equilibrium between native and denatured forms. However, the importance of protein aggregation is increasingly recognized, and it has been suggested that certain aggregated states represent alternative folds of the polypeptide chain. Here, we present a P,T -diagram for myoglobin in which we include the aggregated state and suggest to call it a P,T -state diagram, as not all boundaries are true equilibrium transitions. We observe by Fourier transform infrared spectroscopy that increasing temperature causes the protein to aggregate, but that a subsequent further temperature increase results in the dissociation of this aggregate. Moreover, we observe that moderate pressures stabilize myoglobin against thermal denaturation. We hypothesize that this effect originates from the volume changes associated with the aggregation transition. [source] Influence of an extract from kudzu symbiosomes containing leghemoglobin on in vitro cutaneous procollagen productionINTERNATIONAL JOURNAL OF COSMETIC SCIENCE, Issue 3 2010J. V. Gruber J. Cosmet. Sci., 60, 475,484 (September/October 2009) Synopsis Cytoglobin is a hexacoordinateglobin protein that was recently discovered in mammals. Interestingly, of the four human globin proteins that are now known, hemoglobin, myoglobin, neuroglobin and cytoglobin, the latter appears to have the closest resemblance to strikingly similar proteins expressed in plants. In legumes, these proteins accumulate in symbiosomes (root nodules) of various legumes and are called leghemoglobin. The paper will discuss the ability of an aqueous extract from Pueraria lobata (kudzu) symbiosomes that contains leghemoglobin to stimulate procollagen production in human dermal fibroblasts. This effect may be partly due to the possibility that leghemoglobin may mimic the function of cytoglobin by shuttling oxygen to prolyl-4-hydroxylase, the enzyme responsible for oxidizing proline residues in procollagen bundles. This hypothesis is supported by DNA microarray sequencing data that demonstrate that treatment of normal human dermal fibroblasts (NHDF) with highly purified cytoglobin or leghemoglobin upregulates a number of key collagen-related genes including COL1A1 and COL1A2. [source] Hemoprotein time-resolved X-ray crystallographyIUBMB LIFE, Issue 3 2008Mario Milani Abstract In the last decade the role of structural dynamics in controlling protein function was actively investigated using new and advanced experimental approaches. In particular, time resolved crystallography, despite some practical difficulties, is being used extensively to complement the study of protein structure-function relationships with information on the dynamics, based on experimental evidence. Here we present a short overview of the results obtained on dynamical properties of myoglobin and homologous hemoproteins, where the photosensitive heme-Fe,ligand bond has allowed transient intermediates to be studied by different flash photolysis methods coupled to Laue X-ray diffraction, thus highlighting some of the dynamical events that characterize diffusion of a diatomic ligand to/from the heme in model hemoproteins. © 2008 IUBMB IUBMB Life, 60(3): 154,158, 2008 [source] Histidine and not tyrosine is required for the Peroxide-induced formation of haem to protein cross-linked myoglobinIUBMB LIFE, Issue 8-9 2007Brandon J. Reeder Abstract Peroxide-induced oxidative modifications of haem proteins such as myoglobin and haemoglobin can lead to the formation of a covalent bond between the haem and globin. These haem to protein cross-linked forms of myoglobin and haemoglobin are cytotoxic and have been identified in pathological conditions in vivo. An understanding of the mechanism of haem to protein cross-link formation could provide important information on the mechanisms of the oxidative processes that lead to pathological complications associated with the formation of these altered myoglobins and haemoglobins. We have re-examined the mechanism of the formation of haem to protein cross-link to test the previously reported hypothesis that the haem forms a covalent bond to the protein via the tyrosine 103 residue (Catalano, C. E., Choe, Y. S., Ortiz de Montellano, P. R., J. Biol. Chem. 1989, 10534 - 10541). Comparison of native horse myoglobin, recombinant sperm whale myoglobin and Tyr103 , Phe sperm whale mutant shows that, contrary to the previously proposed mechanism of haem to protein cross-link formation, the absence of tyrosine 103 has no impact on the formation of haem to protein cross-links. In contrast, we have found that engineered myoglobins that lack the distal histidine residue either cannot generate haem to protein cross-links or show greatly suppressed levels of modified protein. Moreover, addition of a distal histidine to myoglobin from Aplysia limacina, that naturally lacks this histidine, restores the haem protein's capacity to generate haem to protein cross-links. The distal histidine is, therefore, vital for the formation of haem to protein cross-link and we explore this outcome. [source] Multiple strategies for O2 transport: from simplicity to complexityIUBMB LIFE, Issue 8-9 2007Paolo Ascenzi Abstract O2carriers (extracellular and intracellular as well as monomeric and multimeric) have evolved over the last billion of years, displaying iron and copper reactive centers; very different O2carriers may co-exist in the same organism. Circulating O2carriers, faced to the external environment, are responsible for maintaining an adequate delivery of O2to tissues and organs almost independently of the environmental O2partial pressure. Then, intracellular globins facilitate O2transfer to mitochondria sustaining cellular respiration. Here, molecular aspects of multiple strategies evolved for O2transport and delivery are examined, from the simplest myoglobin to the most complex giant O2carriers and the red blood cell, mostly focusing on the aspects which have been mainly addressed by the so called 'Rome Group'. [source] Evaluation of stratus® CS Stat fluorimetric analyser for measurement of cardiac markers Troponin I (cTnI), creatine kinase MB (CK-MB), and myoglobinJOURNAL OF CLINICAL LABORATORY ANALYSIS, Issue 6 2001Bénédicte Bénéteau-Burnat Abstract Myoglobin, CK-MB, and Troponin I (cTnI) are cardiac muscle necrosis markers that are useful for detecting acute myocardial infarction (AMI). The Stratus® CS (Dade Behring, Inc.) is a discrete fluorimetric immunoassay analyser designed for the determination of the three cardiac markers from a single sample of whole blood or plasma. Overall analytical performances of the Stratus® CS provided by Dade Behring were evaluated according to the French Society of Clinical Biology guidelines. Within-run imprecision (n = 20) for the three parameters at three levels gave values under 5%, whereas CVs for between-run imprecision (n = 20) were under 6%. The sensitivities were 0.03 ,g/L for cTnI and 0.4 ,g/L for CK-MB. Linearities extended from 0,50 ,g/L for cTnI, 0,140 ,g/L for CK-MB, and 1,900 ,g/L for myoglobin. The results, particularly those obtained on whole-blood samples, correlated well with those obtained on Stratus® II. We did not find any interference with haemolysis, icterus, or lipemia. The system was very easy to use, and fulfills the requirements for the analysis of the three cardiac markers in patients with acute chest pain in emergency situations. J. Clin. Lab. Anal. 15:314,318, 2001. © 2001 Wiley-Liss, Inc. [source] Is skeletal muscle damaged by the oxidative stress following anaerobic exercise?JOURNAL OF CLINICAL LABORATORY ANALYSIS, Issue 5 2001Hiroshi Ihara Abstract We investigated whether the injury of skeletal muscle owing to the action of free radicals and the subsequent oxidative damage to tissues occurred during anaerobic exercise. To estimate injury to skeletal muscle, we determined certain indices of oxidative damage to skeletal muscle; i.e., leukocyte counts, concentrations of hypoxanthine, xanthine, urate, tissue- and serum-type CK-M isoforms, myoglobin, and total antioxidant capacity (TAC) of serum. Blood for these tests was collected at 3 min post-exercise. Post-anaerobic exercise concentrations of lactate were significantly increased from pre-exercise. The neutrophil and lymphocyte counts and alanine concentration were significantly increased by anaerobic exercise, even when the results were corrected for plasma volume changes; the plasma concentrations of hypoxanthine, urate, and TAC of serum were also significantly increased. The plasma concentration of xanthine was negatively correlated with TAC of serum. The activities of tissue- and serum-type CK-M were significantly increased post-exercise. When the hypoxanthine, urate, TAC of serum, myoglobin, and tissue- and serum-type CK-M were corrected for plasma volume changes, the post-exercise increases were no longer significantly different from the pre-exercise results. We suggest that these latter test results following anaerobic exercise exclude the presence of oxidative damage to skeletal muscle. J. Clin. Lab. Anal. 15:239,243, 2001. © 2001 Wiley-Liss, Inc. [source] Species-Specific Effects of Sarcoplasmic Extracts on Lipid Oxidation in vitroJOURNAL OF FOOD SCIENCE, Issue 1 2009R. Ramanathan ABSTRACT:, The degree to which lipid and myoglobin (Mb) oxidation processes interact in meat can be species-specific. We investigated the effects of beef and pork sarcoplasmic extracts containing different Mb concentrations on lipid oxidation in a liposome system. Sarcoplasm was extracted from beef and pork longissimus dorsi and psoas major muscles. Beef sarcoplasm was diluted with 0.1 M phosphate buffer to obtain a Mb concentration equivalent to that in pork sarcoplasm. Conversely, equine heart Mb was added to pork sarcoplasm to match the myoglobin concentration of beef sarcoplasm. This resulted in beef and pork sarcoplasms, each with 2 different Mb concentrations for the longissimus (0.02 mM and 0.07 mM) and psoas (0.05 and 0.12 mM). Sarcoplasm (or phosphate buffer control) was incorporated within a phosphatidylcholine liposome preparation and incubated at 25°C. Thiobarbituric acid reactive substances (TBARS) were measured at 0, 30, 60, 90, and 120 min of incubation. Regardless of species, greater Mb concentration within the sarcoplasm increased lipid oxidation (P < 0.05). Across muscles, pork sarcoplasm had lower TBARS values than beef sarcoplasm (P < 0.05). Our results suggest that pork sarcoplasm has a lesser effect on lipid oxidation than beef sarcoplasm for a common Mb concentration. However, increased myoglobin concentration within sarcoplasm promotes lipid oxidation regardless of species. [source] Influence of Cooking Rate, Endpoint Temperature, Post-cook Hold Time, and Myoglobin Redox State on Internal Color Development of Cooked Ground Beef PattiesJOURNAL OF FOOD SCIENCE, Issue 3 2006Suzanne M. Ryan ABSTRACT: Three experiments investigated cooking rate, endpoint temperature, post-cook holding time, and raw myoglobin redox-state effects on ground beef internal cooked color. In Experiment 1, patties were cooked to endpoint temperatures of 65.6°C, 71.1°C, 76.7°C, 82.2°C, or 87.8°C rapidly (1°C/s), slowly (0.2°C/s), or rapidly with 6-min post-cook holding time at 104°C. Patties cooked slowly to less than 76.7°C were more well done (P < 0.05) in appearance than those cooked rapidly. Rapidly-cooked patties cooked to less than 82.2°C and held for 6 min after cooking had less pinkness, more myoglobin denaturation, and a more well-done appearance than did rapidly cooked patties with no holding time (P < 0.05). In Experiment 2, increasing post-cook holding time (1, 3, 6, or 12 min) after rapid cooking to 71.1°C, 76.7°C, or 82.2°C decreased pinkness and increased myoglobin denaturation (P < 0.05), with no benefit beyond 6 min (P > 0.05). In Experiment 3, patties cooked rapidly to 71.1°C, 76.7°C, or 82.2°C from a predominantly raw oxymyoglobin state were less pink and had more denatured myoglobin than did those cooked from a predominantly deoxymyoglobin state (P < 0.05). Prediction equations determined that 80% of myoglobin must be denatured to create a well-done appearance. Using a slow cooking rate, post-cook holding time, or cooking from a highly oxygenated state will increase myoglobin denaturation and foster a well-done appearance. [source] Internal Premature Browning in Cooked Steaks from Enhanced Beef Round Muscles Packaged in High-oxygen and Ultra-low Oxygen Modified AtmospheresJOURNAL OF FOOD SCIENCE, Issue 2 2004M. SEYFERT ABSTRACT: Beef round muscles were injection-enhanced to 6%, packaged in high-oxygen (HiOx) or ultra-low oxygen (LoOx) modified atmospheres, stored 7 d and displayed 2 d (HiOx) or stored 16 d and displayed 1 d (LoOx) at 0 °C, and cooked to 71.1 °C. Raw internal color for steaks in HiOx was lighter, redder, more yellow and saturated, and had more oxymyoglobin and less deoxymyoglobin than steaks in LoOx (P < 0.0001). Cooked internal color of steaks from HiOx appeared prematurely brown and was darker, less red, yellow, and saturated, and had more denatured myoglobin than steaks from LoOx (P < 0.0001). This study presents conclusive evidence that modified-atmosphere packaging influences internal cooked color development of beef steaks. [source] | |||||||||||||||||||||||||||||||||||||||||||