Cloning, sequence analysis, and expression in Escherichia coli of a streptococcal plasmin receptor

R. Lottenberg, C. C. Broder, M. D.P. Boyle, S. J. Kain, B. L. Schroeder, R. Curtiss

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Abstract

Plasmin(ogen) receptors are expressed by many gram-positive and gram- negative bacteria. We previously isolated a plasmin receptor from a pathogenic group A streptococcal strain (C. C. Broder, R. Lottenberg, G. O. von Mering, K. H. Johnston, and M. D. P. Boyle, J. Biol. Chem. 266:4922-4928, 1991). The gene encoding this plasmin receptor, plr, was isolated from a λ gt11 library of chromosomal DNA from group A streptococcal strain 64/14 by screening plaques with antibodies raised against the purified streptococcal plasmin receptor protein. The gene was subcloned by using a low-copy-number plasmid and stably expressed in Escherichia coli, resulting in the production of an immunoreactive and functional receptor protein. The DNA sequence of the gene contained an open reading frame encoding 335 amino acids with a predicted molecular weight of 35,787. Upstream of the open reading frame, putative promoter and ribosomal binding site sequences were identified. The experimentally derived amino acid sequences of the N terminus and three cyanogen bromide fragments of the purified streptococcal plasmin receptor protein corresponded to the predicted sequence encoded by plr. The deduced amino acid sequence for the plasmin receptor protein revealed significant similarity (39 to 54% identical amino acid residues) to glyceraldehyde 3- phosphate dehydrogenases.

Original languageEnglish (US)
Pages (from-to)5204-5210
Number of pages7
JournalJournal of bacteriology
Volume174
Issue number16
DOIs
StatePublished - Jan 1 1992

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ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

Cite this

Lottenberg, R., Broder, C. C., Boyle, M. D. P., Kain, S. J., Schroeder, B. L., & Curtiss, R. (1992). Cloning, sequence analysis, and expression in Escherichia coli of a streptococcal plasmin receptor. Journal of bacteriology, 174(16), 5204-5210. https://doi.org/10.1128/jb.174.16.5204-5210.1992