Document Detail

Proteomics-based identification of outer-membrane proteins responsible for import of macromolecules in Sphingomonas sp. A1: alginate-binding flagellin on the cell surface.
MedLine Citation:
PMID:  16229468     Owner:  NLM     Status:  MEDLINE    
A nonmotile gram-negative bacterium, Sphingomonas sp. A1, directly incorporates macromolecules such as alginate through a "super-channel" consisting of a pit formed on the cell surface, alginate-binding proteins in the periplasm, and an ATP-binding cassette transporter in the inner membrane. Here, we demonstrate the proteomics-based identification of cell-surface proteins involved in the formation of the pit and/or import of alginate. Cell-surface proteins were prepared from the outer membrane released as vesicles during the conversion of intact cells to spheroplasts. Seven proteins (p1-p7) with acidic isoelectric points were inducibly expressed in the outer membrane of strain A1 cells grown on alginate and showed significant identity with bacterial cell-surface proteins (p1-p4, TonB-dependent outer-membrane transporter; p5 and p6, flagellin; and p7, lipoprotein). Each mutant with a disruption of the p1-p4 or p6 gene showed significant growth retardation in the alginate medium. Flagellin homologues (p5 and p6) were further analyzed because strain A1 forms no flagellum. p5 was found to be uniformly distributed on the cell surface by immunogold-labeling electron microscopy and to exhibit alginate binding with a nanomolar dissociation constant by a surface plasmon resonance sensor. The cell surface of the p6 gene disruptant differed from that of the wild-type strain A1 in that pit formation was incomplete and cell-surface structures shifted from pleats to networks. These results suggest that, distinct from bacterial flagellins constituting a helical filament of flagella, strain A1 cell-surface flagellin homologues function as receptors for alginate and/or regulators of cell-surface structures.
Wataru Hashimoto; Jinshan He; Yushin Wada; Hirokazu Nankai; Bunzo Mikami; Kousaku Murata
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Biochemistry     Volume:  44     ISSN:  0006-2960     ISO Abbreviation:  Biochemistry     Publication Date:  2005 Oct 
Date Detail:
Created Date:  2005-10-18     Completed Date:  2006-04-03     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0370623     Medline TA:  Biochemistry     Country:  United States    
Other Details:
Languages:  eng     Pagination:  13783-94     Citation Subset:  IM    
Laboratory of Basic and Applied Molecular Biotechnology, Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Uji, Kyoto 611-0011, Japan.
Data Bank Information
Bank Name/Acc. No.:
GENBANK/AB183457;  AB183458;  AB211539;  AB211540;  AB211541;  AB211542;  AB211543
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MeSH Terms
Alginates / metabolism*
Amino Acid Sequence
Bacterial Outer Membrane Proteins / chemistry,  metabolism*
Electrophoresis, Polyacrylamide Gel
Flagellin / genetics,  metabolism*
Glucuronic Acid / metabolism
Hexuronic Acids / metabolism
Microscopy, Electron
Molecular Sequence Data
Protein Binding
Sphingomonas / chemistry,  metabolism*
Reg. No./Substance:
0/Alginates; 0/Bacterial Outer Membrane Proteins; 0/Hexuronic Acids; 12777-81-0/Flagellin; 576-37-4/Glucuronic Acid; 9005-32-7/alginic acid

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