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Numerous Proteins (numerous + protein)
Selected AbstractsSodium thiosulfate as an effective antioxidant substance at experimental mycotoxin zearalenon poisoningJOURNAL OF NEUROCHEMISTRY, Issue 2002M. K. Karagyozyan Mycotoxin zearalenon (MZ) in concentration 2.5,15.0 ,g/mL of alcohol solution activates the reactions of monoamines biosynthesis, while 20.0,25.0 ,g/mL of MZ has a contrarary effect. The molecular mechanism of changes noticed is conditioned by action of MZ on numerous proteins functioning in chromaffin granules mainly in their membranes, such as the cytochrome B561, an acidic cooper-containing protein and the membrane-bound form of dopamine-,-monooxigenase, which catalyses the reaction of transformation of dopamine into noradrenaline. It has been established also that in the brain mitochondrial and microsomal membranes the MZ induces pronounced abnormalities in the ratio of phospholipid-phospholipid interrelations. These changes are conditioned by significant intensification of the phospholipase A2 and phosphatidylserine (PS) decarboxylase activites, with formation of high concentrations of lysophosphatidylcholines, free polyenic fatty acids, lipid peroxides and by increasing of PS quantity in the systems studied. Using on this background single intramuscular administration of 1.0 mL of 10.0% aquaus solution of sodium thiosulfate (ST) normalizes and establishes the initial level of phospholipid (PL) metabolism intensity. The content of PL in the investigated membranes remains unchanged if ST was administrated before the MZ poisoning modulation. Antioxidant properties of ST are conditioned in particular by elevation of PS quantity, which are of great importance in stimulation of cell respiratory function, hence the cell activity in general. [source] Identification, molecular cloning, and cellular distribution of the rat homolog of minichromosome maintenance protein 7 (MCM7) in the rat testis,MOLECULAR REPRODUCTION & DEVELOPMENT, Issue 7 2006Emmanuelle Com Abstract As part of a program to decipher the rat testicular proteome, we studied spermatogonia and identified numerous proteins including the human homolog of the Minichromosome Maintenance Protein 7 (MCM7). MCM7 has been implicated in DNA replication in various species, but had not been detected in the testis. Here we describe the cellular distribution of MCM7 transcripts and protein, and their testicular ontogenetic expression. The full-length coding region of the rat MCM7 was also characterized. Northern blot analyses showed that MCM7 transcripts are more abundant in the testis than other organs and confirmed the presence of the 2.4 kb MCM7 transcript at all ages studied. Interestingly, two additional transcripts of 3.2 and 1.6 kb were found from 26 days post partum onwards, when spermatocytes and spermatids accumulate within the tubules. This was confirmed in isolated cell types: the three MCM7 transcripts were observed in meiotic and post-meiotic germ cells. The 3.2 kb isoform has an extended 5, untranslated region (UTR) and the 1.6 kb transcript is the result of alternative splicing of five exons. Western blot and immunohistochemistry experiments evidenced abundant MCM7 in proliferating gonocytes and Sertoli cells in the fetal testis. In the adult testis, an intense signal was observed in spermatogonia and primary spermatocytes. We conclude that the Mcm7 is one example of genes that are differently transcribed and translated in somatic and spermatogenetic cells in mammals. Further work is required to determine the roles of MCM7 in spermatogonia and germ lineage. Mol. Reprod. Dev. © 2006 Wiley-Liss, Inc. [source] Characterisation of Plasmodium invasive organelles; an ookinete microneme proteomePROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 5 2009Kalpana Lal Dr. Abstract Secretion of microneme proteins is essential to Plasmodium invasion but the molecular composition of these secretory organelles remains poorly defined. Here, we describe the first Plasmodium microneme proteome. Purification of micronemes by subcellular fractionation from cultured ookinetes was confirmed by enrichment of known micronemal proteins and electron microscopy. Quantitation of electron micrographs showed >14-fold microneme enrichment compared to the intact ookinete, such that micronemes comprised 85% of the identifiable organelles in the fraction. Gel LC-MS/MS of the most abundant protein constituents of the fraction identified three known micronemal proteins chitinase, CTRP, SOAP, together with protein disulphide isomerase (PDI) and HSP70. Highly sensitive MudPIT shotgun proteomics described a total of 345 proteins in the fraction. M1 aminopeptidase and PDI, the former a recognised target of drug development, were both shown to have a micronemal location by IFA. We further identified numerous proteins with established vesicle trafficking and signaling functions consistent with micronemes being part of a regulated secretory pathway. Previously uncharacterised proteins comprise the largest functional group of the microneme proteome and will include secreted proteins important to invasion. [source] Flow cytometry-assisted purification and proteomic analysis of the corticotropes dense-core secretory granulesPROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 18 2008Daniel J. Gauthier Abstract The field of organellar proteomics has emerged as an attempt to minimize the complexity of the proteomics data obtained from whole cell and tissue extracts while maximizing the resolution on the protein composition of a single subcellular compartment. Standard methods involve lengthy density-based gradient and/or immunoaffinity purification steps followed by extraction, 1-DE or 2-DE, gel staining, in-gel tryptic digestion, and protein identification by MS. In this paper, we present an alternate approach to purify subcellular organelles containing a fluorescent reporter molecule. The gel-free procedure involves fluorescence-assisted sorting of the secretory granules followed by gentle extraction in a buffer compatible with tryptic digestion and MS. Once the subcellular organelle labeled, this procedure can be done in a single day, requires no major modification to any instrumentation and can be readily adapted to the study of other organelles. When applied to corticotrope secretory granules, it led to a much enriched granular fraction from which numerous proteins could be identified through MS. [source] Proteome analysis of the phenotypic variation process in Photorhabdus luminescensPROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 9 2006Evelyne Turlin Dr. Abstract Photorhabdus luminescens is an insect pathogen associated with specific soil nematodes. The bacterium has a complex life cycle with a symbiotic stage in which bacteria colonize the intestinal tract of the nematodes, and a pathogenic stage against susceptible larval-stage insect. Symbiosis-"deficient" phenotypic variants (known as secondary forms) arise during prolonged incubation. Correspondence analysis of the in silico proteome translated from the genome sequence of strain TT01 identified two major biases in the amino acid composition of the proteins. We analyzed the proteome, separating three classes of extracts: cellular, extracellular, and membrane-associated proteins, resolved by 2-DE. Approximately 450 spots matching the translation products of 231 different coding DNA sequences were identified by PMF. A comparative analysis was performed to characterize the protein content of both variants. Differences were evident during stationary growth phase. Very few proteins were found in variant II supernatants, and numerous proteins were lacking in the membrane-associated fraction. Proteins up-regulated by the phenotypic variation phenomenon were involved in oxidative stress, energy metabolism, and translation. The transport and binding of iron, sugars and amino acids were also affected and molecular chaperones were strongly down-regulated. A potential role for H-NS in phenotypic variation control is discussed. [source] Versatile protein microarray based on carbohydrate-binding modulesPROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 7 2005Keren Ofir Abstract Non-DNA microarrays, such as protein, peptide and small molecule microarrays, can potentially revolutionize the high-throughput screening tools currently used in basic and pharmaceutical research. However, fundamental obstacles remain that limit their rapid and widespread implementation as an alternative bioanalytical approach. These include the prerequisite for numerous proteins in active and purified form, ineffectual immobilization strategies and inadequate means for quality control of the considerable numbers of multiple reagents. This study describes a simple yet efficient strategy for the production of non-DNA microarrays, based on the tenacious affinity of a carbohydrate-binding module (CBM) for its three-dimensional substrate, i.e., cellulose. Various microarray formats are described, e.g., conventional and single-chain antibody microarrays and peptide microarrays for serodiagnosis of human immunodeficiency virus patients. CBM-based microarray technology overcomes many of the previous obstacles that have hindered fabrication of non-DNA microarrays and provides a technically simple but effective alternative to conventional microarray technology. [source] Prostate cancer expression of runt-domain transcription factor Runx2, a key regulator of osteoblast differentiation and functionTHE PROSTATE, Issue 1 2003Kristen D. Brubaker Abstract BACKGROUND Prostate cancer (CaP) bone metastases express numerous proteins associated with bone cells. Specific transcription factors, including Runx2, regulate the expression of many bone-related factors in osteoblasts. Expression of these transcription factors in CaP may be linked to the ability of CaP bone metastases to influence bone remodeling. METHODS CaP tissues and cell lines were analyzed for expression of Runx2 mRNA by RT-PCR and in situ hybridization, and protein by immunohistochemistry, Western blotting, and electrophoretic mobility shift assays (EMSA). RESULTS Runx2 mRNA and protein were detected in CaP tissues and cell lines. A specific Runx2: OSE2 complex could be formed with PC-3 nuclear extracts. CONCLUSIONS Expression of Runx2 in CaP may be the molecular switch that is associated with expression of various bone-specific factors in CaP. In turn, expression of these factors can influence bone remodeling and possibly play a role in the growth and survival of CaP in bone. Prostate 56: 13,22, 2003. © 2003 Wiley-Liss, Inc. [source] | ||||||||||