Fine tangled pili expressed by Haemophilus ducreyi are a novel class of pili

被引:60
作者
Brentjens, RJ
Ketterer, M
Apicella, MA
Spinola, SM
机构
[1] INDIANA UNIV, SCH MED, DEPT MED, INDIANAPOLIS, IN 46202 USA
[2] INDIANA UNIV, SCH MED, DEPT MICROBIOL, INDIANAPOLIS, IN 46202 USA
[3] INDIANA UNIV, SCH MED, DEPT IMMUNOL, INDIANAPOLIS, IN 46202 USA
[4] SUNY BUFFALO, SCH MED, DEPT MICROBIOL, BUFFALO, NY 14214 USA
[5] UNIV IOWA, COLL MED, DEPT MICROBIOL, IOWA CITY, IA 52242 USA
关键词
D O I
10.1128/jb.178.3.808-816.1996
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Haemophilus ducreyi synthesizes fine, tangled pill composed predominantly of a protein whose apparent molecular weight is 24,000 (24K). A hybridoma, 2D8, produced a monoclonal antibody (MAb) that bound to a 24K protein in H. ducreyi strains isolated from diverse geographic locations. A lambda gt11 H. ducreyi library was screened with MAb 2D8. A 3.5-kb chromosomal insert from one reactive plaque was amplified and ligated into the pCRII vector. The recombinant plasmid, designated pHD24, expressed a 24K protein in Escherichia coli INV alpha F' that bound MAb 2D8. The coding sequence of the 24K gene was localized by exonuclease III digestion. The insert contained a 570-bp open reading frame, designated ftpA (fine, tangled pill). Translation of ftpA predicted a polypeptide with a molecular weight of 21.1K. The predicted N-terminal amino acid sequence of the polypeptide encoded by ftpA was identical to the N-terminal amino acid sequence of purified pilin and lacked a cleavable signal sequence. Primer extension analysis of ftpA confirmed the lack of a leader peptide. The predicted amino acid sequence lacked homology to known pilin sequences but shared homology,vith the sequences of E. coli Dps and Treponema pallidum antigen TpF1 or 4D, proteins which associate to form ordered rings. An isogenic pilin mutant, H. ducreyi 35000ftpA::mTn3(Cm), was constructed by shuttle mutagenesis and did not contain pill when examined by electron microscopy. We conclude that H. ducreyi synthesizes fine, tangled pill that are composed of a unique major subunit, which may be exported by a signal sequence independent mechanism.
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页码:808 / 816
页数:9
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