SUBCELLULAR-LOCALIZATION OF CLOSTRIDIUM-THERMOCELLUM ORF3P, A PROTEIN CARRYING A RECEPTOR FOR THE DOCKING SEQUENCE BORNE BY THE CATALYTIC COMPONENTS OF THE CELLULOSOME

被引:66
作者
SALAMITOU, S
LEMAIRE, M
FUJINO, T
OHAYON, H
GOUNON, P
BEGUIN, P
AUBERT, JP
机构
[1] INST PASTEUR, DEPT BIOTECHNOL, UNITE PHYSIOL CELLULAIRE, F-75724 PARIS 15, FRANCE
[2] INST PASTEUR, DEPT BIOTECHNOL, CNRS, URA 1300, F-75724 PARIS 15, FRANCE
[3] INST PASTEUR, MICROSCOPIE ELECTR STN, F-75724 PARIS 15, FRANCE
关键词
D O I
10.1128/JB.176.10.2828-2834.1994
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The ORF3 gene of Clostridium thermocellum encodes a polypeptide (ORF3p) which contains a receptor domain for the docking sequence borne by the catalytic subunits of the cellulosome and a triplicated domain related to some bacterial cell surface proteins. It was thus surmised that ORF3p is a surface protein. In this study, this hypothesis was confirmed. Subcellular fractionation, Western blotting (immunoblotting), and electron microscopy of immunocytochemically labeled cells indicated that ORF3p produced by C. thermocellum was located in the outer surface layer of the bacterium. This layer appeared to consist of a soft matrix shedding off particulate fragments, Nonsedimenting ORF3p derived from sonicated cells was associated with high-molecular-mass fractions (>20 MDa), probably corresponding to fragments of the outer cell layer. The same high-molecular-mass fractions also contained the cellulosomal marker CipA. Contrary to CipA, however, ORF3p was not associated with 2- to 4-MDa fractions corresponding to individual cellulosomes, and a significant fraction of ORF3p failed to bind to cellulose. It is proposed that ORF3 and ORF3p be renamed olpA and OlpA, respectively (for outer layer protein).
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页码:2828 / 2834
页数:7
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