| A new photoprobe for studying biological activities of secreted phospholipases A2. | |
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MedLine Citation:
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PMID: 17303214 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Ammodytoxin (Atx) is a snake venom phospholipase A2 (sPLA2s) with presynaptic toxicity, anticoagulant activity and the ability to influence cell cycle progression. These multiple physiological activities make this molecule a promising tool for studying processes influenced by the highly homologous mammalian sPLA2s-for example cell proliferation and apoptosis. Secreted PLA2s can act on cells as enzymes or as ligands for cellular receptors. To further characterize the sPLA2-binding molecules in cells we have developed a new method based on AtxC and a biotin-containing cross-linking reagent sulfo-SBED which possesses both an amine-reactive and a photo-reactive site, together with a biotin moiety that enables specific detection and affinity-based concentration. The biological activity of the AtxC derivatized by sulfo-SBED was demonstrated by biotin-tagging of calmodulin and R25, both known AtxC targets, but not of other proteins. In addition, using the new protocol we specifically labelled 14-3-3 proteins, protein disulfide isomerase and two unknown proteins of 45 and 46kDa in the mitochondrial-synaptosomal fraction of porcine cerebral cortex, none of which could be tagged by the previously used methods. The new methodology, which can be used for any sPLA2, constitutes a novel approach to discovering and purifying sPLA2-binding proteins, to studying the topology of their respective complexes and to following sPLA2s in different biological systems. |
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Authors:
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Lidija Kovacic; Jernej Sribar; Igor Krizaj |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2007-02-15 |
Journal Detail:
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Title: Bioorganic chemistry Volume: 35 ISSN: 0045-2068 ISO Abbreviation: Bioorg. Chem. Publication Date: 2007 Aug |
Date Detail:
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Created Date: 2007-07-03 Completed Date: 2007-09-05 Revised Date: 2007-11-15 |
Medline Journal Info:
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Nlm Unique ID: 1303703 Medline TA: Bioorg Chem Country: United States |
Other Details:
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Languages: eng Pagination: 295-305 Citation Subset: IM |
Affiliation:
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Department of Biochemistry and Molecular Biology, Jozef Stefan Institute, Jamova 39, SI-1000 Ljubljana, Slovenia. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Biotin Cross-Linking Reagents Humans Molecular Probe Techniques* Phospholipases A / analysis, metabolism*, secretion Phospholipases A2 Photochemistry Protein Binding Protein Interaction Mapping / methods* Viper Venoms / diagnostic use* |
| Chemical | |
Reg. No./Substance:
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0/Cross-Linking Reagents; 0/Viper Venoms; 58-85-5/Biotin; 97089-72-0/ammodytoxin A; EC 3.1.1.-/Phospholipases A; EC 3.1.1.4/Phospholipases A2 |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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