| The surface location of individual residues in a bacterial S-layer protein. | |
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MedLine Citation:
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PMID: 18262545 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Bacterial surface layer (S-layer) proteins self-assemble into large two-dimensional crystalline lattices that form the outermost cell-wall component of all archaea and many eubacteria. Despite being a large class of self-assembling proteins, little is known about their molecular architecture. We investigated the S-layer protein SbsB from Geobacillus stearothermophilus PV72/p2 to identify residues located at the subunit-subunit interface and to determine the S-layer's topology. Twenty-three single cysteine mutants, which were previously mapped to the surface of the SbsB monomer, were subjected to a cross-linking screen using the photoactivatable, sulfhydryl-reactive reagent N-[4-(p-azidosalicylamido)butyl]-3'-(2'-pyridyldithio)propionamide. Gel electrophoretic analysis on the formation of cross-linked dimers indicated that 8 out of the 23 residues were located at the interface. In combination with surface accessibility data for the assembled protein, 10 residues were assigned to positions at the inner, cell-wall-facing lattice surface, while 5 residues were mapped to the outer, ambient-exposed lattice surface. In addition, the cross-linking screen identified six positions of intramolecular cross-linking within the assembled protein but not in the monomeric S-layer protein. Most likely, these intramolecular cross-links result from conformational changes upon self-assembly. The results are an important step toward the further structural elucidation of the S-layer protein via, for example, X-ray crystallography and cryo-electron microscopy. Our approach of identifying the surface location of residues is relevant to other planar supramolecular protein assemblies. |
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Authors:
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Helen Kinns; Stefan Howorka |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2008-01-16 |
Journal Detail:
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Title: Journal of molecular biology Volume: 377 ISSN: 1089-8638 ISO Abbreviation: J. Mol. Biol. Publication Date: 2008 Mar |
Date Detail:
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Created Date: 2008-03-10 Completed Date: 2008-03-28 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 2985088R Medline TA: J Mol Biol Country: England |
Other Details:
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Languages: eng Pagination: 589-604 Citation Subset: IM |
Affiliation:
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Department of Chemistry, University College London, Christopher Ingold Building, 20 Gordon Street, London WC1H 0AJ, England, UK. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Bacillaceae
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chemistry*,
metabolism Bacterial Proteins / chemistry*, isolation & purification, metabolism*, ultrastructure Cross-Linking Reagents / chemistry Cysteine / genetics, metabolism Diphosphonates / chemistry Gene Expression Membrane Proteins / chemistry*, isolation & purification, metabolism*, ultrastructure Microscopy, Electron, Transmission Mutation / genetics Recombinant Proteins / chemistry, genetics, isolation & purification, metabolism |
| Grant Support | |
ID/Acronym/Agency:
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//Biotechnology and Biological Sciences Research Council |
| Chemical | |
Reg. No./Substance:
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0/Bacterial Proteins; 0/Cross-Linking Reagents; 0/Diphosphonates; 0/Membrane Proteins; 0/Recombinant Proteins; 52-90-4/Cysteine; 56269-44-4/azacycloheptane-2,2-diphosphonate |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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