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Peptide Degradation (peptide + degradation)
Selected AbstractsProfiling of neuropeptides released at the stomatogastric ganglion of the crab, Cancer borealis with mass spectrometryJOURNAL OF NEUROCHEMISTRY, Issue 1 2005Cyrus P. Billimoria Abstract Studies of release under physiological conditions provide more direct data about the identity of neuromodulatory signaling molecules than studies of tissue localization that cannot distinguish between processing precursors and biologically active neuropeptides. We have identified neuropeptides released by electrical stimulation of nerves that contain the axons of the modulatory projection neurons to the stomatogastric ganglion of the crab, Cancer borealis. Preparations were bathed in saline containing a cocktail of peptidase inhibitors to minimize peptide degradation. Both electrical stimulation of projection nerves and depolarization with high K+ saline were used to evoke release. Releasates were desalted and then identified by mass using MALDI,TOF (matrix-assisted laser desorption/ionization,time-of-flight) mass spectrometry. Both previously known and novel peptides were detected. Subsequent to electrical stimulation proctolin, Cancer borealis tachykinin-related peptide (CabTRP), FVNSRYa, carcinustatin-8, allatostatin-3 (AST-3), red pigment concentrating hormone, NRNFLRFa, AST-5, SGFYANRYa, TNRNFLRFa, AST-9, orcomyotropin-related peptide, corazonin, Ala13-orcokinin, and Ser9-Val13-orcokinin were detected. Some of these were also detected after high K+ depolarization. Release was calcium dependent. In summary, we have shown release of the neuropeptides thought to play an important neuromodulatory role in the stomatogastric ganglion, as well as numerous other candidate neuromodulators that remain to be identified. [source] Elimination and exchange of trifluoroacetate counter-ion from cationic peptides: a critical evaluation of different approachesJOURNAL OF PEPTIDE SCIENCE, Issue 3 2008Stéphane Roux Abstract Most synthesized peptides are nowadays produced using solid-phase procedures. Due to cleavage and purification conditions, they are mainly obtained in the presence of trifluoroacetic acid (TFA) and, for cationic peptides, as trifluoroacetate (TF-acetate) salts. However, TF-acetate interferes with physicochemical characterizations using infrared spectroscopy and might significantly affect the in vivo studies. Thus, TF-acetate exchange by another counter-ion is often required. Up to now, the classical procedure has consisted of freeze-drying the peptide several times in the presence of an excess of a stronger acid than TFA (pKa ,0): generally HCl (pKa = , 7). This approach means that working at pH < 1 can induce peptide degradation. We therefore tested three different approaches to exchange the tightly bound TF-acetate counter-ion from the dicationic octapeptide lanreotide: (i) reverse-phase HPLC, (ii) ion-exchange resin, and (iii) deprotonation/reprotonation cycle of the amino groups. The first two approaches allow the partial to almost complete exchange of the TF-acetate counter-ion by another ion from an acid weaker than TFA, such as acetic acid (pKa = 4.5), and the third requires a basic solution that permits the complete removal of TF-acetate counter-ion. The efficiency of these three procedures was tested and compared by using different analytical techniques such as 19F-NMR, 1H-NMR and attenuated total reflectance Fourier transformed infrared spectroscopy (ATR FT-IR). We also show that ATR-IR can be used to monitor the TFA removal. The counter-ion exchange procedures described in this study are easy to carry out, fast, harmless and reproducible. Moreover, two of them offer the very interesting possibility of exchanging the initial TF-acetate by any other counter-ion. Copyright © 2007 European Peptide Society and John Wiley & Sons, Ltd. [source] Synthesis of the C -terminal domain of the tissue inhibitor of metalloproteinases-1(TIMP-1)JOURNAL OF PEPTIDE SCIENCE, Issue 7 2003József Bódi Abstract According to recent investigations, the C -terminal domain of the tissue inhibitor of matrix metalloproteinases-1 (TIMP-1) is responsible for some biological effects that are independent of the enzyme-inhibiting effect of the N -terminal domain of the molecule. The C -terminal domain has been prepared for structure,biological activity investigations. After the chemical synthesis and the folding of the linear peptide, LC-MS and MALDI-MS analysis revealed that two isomers with different disulphide bond arrangements were formed. Since more than 30 folding experiments resulted in products with a very similar HPLC-profile, it was concluded that in the absence of the TIMP-1 N -terminal domain no entirely correct folding of the C -terminal domain occurred. Furthermore, it was observed that, in spite of several purification steps, mercury(II) ions were bound to the 6SH-linear peptide; it was demonstrated,using disulphide bonded TIMP-1(Cys145 -Cys166) as a model,that mercury(II) ions can cause peptide degradation at pH 7.8 as well as in 0.1% trifluoroacetic acid. Copyright © 2003 European Peptide Society and John Wiley & Sons, Ltd. [source] Functionality of lactic acid bacteria peptidase activities in the hydrolysis of gliadin-like fragmentsLETTERS IN APPLIED MICROBIOLOGY, Issue 5 2008C.L. Gerez Abstract Aims:, To evaluate the role of the peptidase activities from sourdough lactic acid bacteria (LAB) in the degradation of ,-gliadin fragments. Methods and Results:, Different proline-containing substrates were hydrolysed by LAB indicating pro-specific peptidase activities. Lactobacillus plantarum CRL 775 and Pediococcus pentosaceus CRL 792 displayed the highest tri- and di-peptidase activities, respectively. Lactobacillus plantarum strains hydrolysed more than 60%,-gliadin fragments corresponding to the 31,43 and 62,75 amino acids in the protein after 2 h. None of the LAB strains alone could hydrolyse 57,89 ,-gliadin peptide; however, the combination of L. plantarum CRL 775 and P. pentosaceus CRL 792 led to hydrolysis (57%) of this peptide in 8 h. Conclusions:, The capacity of LAB strains to degrade ,-gliadin fragments was not correlated to individual peptidase activities. Several strains separately degraded the 31,43 and 62,75 ,-gliadin fragments, while the 57,89 peptide degradation was associated with the combination of peptidase profiles from pooled LAB strains. This is the first report on the peptide hydrolase system of sourdough pediococci and its ability to reduce ,-gliadin fragments. Significance and Impact of the Study:, This study contributes to a better knowledge of sourdough LAB proteolytic system and its role in the degradation of proline-rich ,-gliadin peptides involved in celiac disease. [source] Differences between the abilities of tegaserod and motilin receptor agonists to stimulate gastric motility in vitroBRITISH JOURNAL OF PHARMACOLOGY, Issue 4 2007E M Jarvie Background and purpose: Motilin or 5-HT4 receptor agonists stimulate gastrointestinal motility. Differences in activity are suggested but direct comparisons are few. A method was devised to directly compare the gastric prokinetic activities of motilin, the motilin receptor agonist, erythromycin, and the 5-HT4 receptor agonist, tegaserod. Experimental approach: Gastric prokinetic-like activity was assessed by measuring the ability to facilitate cholinergically-mediated contractions evoked by electrical field stimulation (EFS) in rabbit isolated stomach. Comparisons were made between potency, maximal activity and duration of responses. Key results: Rabbit motilin (r.motilin) 0.003,0.3,M, [Nle13]motilin 0.003,0.3,M, erythromycin 0.3,10,M and tegaserod 0.1,10,M caused concentration , dependent potentiation of EFS-evoked contractions. The potency ranking was r.motilin = [Nle13]motilin > tegaserod > erythromycin. The Emax ranking was r.motilin = [Nle13]motilin = erythromycin > tegaserod. Responses to r.motilin and [Nle13]motilin faded rapidly (t1/2 9 and 11 min, respectively) whereas those to erythromycin and tegaserod were maintained longer (t1/2 24 and 28 min). The difference did not appear to be due to peptide degradation. A second application of [Nle13]motilin was excitatory after 60 min contact and fade of the initial response (responses to 0.03 and 0.1,M [Nle13]motilin were not different from those caused by the first application). Conclusions and implications: Prokinetic-like activities of the 5-HT4 agonist tegaserod and the motilin receptor agonists were compared by measuring changes in cholinergically-mediated contractions. This novel approach highlighted important differences between classes (greater Emax of motilin, compared with tegaserod) and for the first time, within each class (short t1/2 for motilin, compared with erythromycin). British Journal of Pharmacology (2007) 150, 455,462. doi:10.1038/sj.bjp.0707118 [source] | ||||||||||