Cloning, sequence analysis, and expression in Escherichia coli of a streptococcal plasmin receptor.
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 lambda 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.
Lottenberg
R
R
Department of Medicine, University of Florida, Gainesville 32610.
Broder
C C
CC
Boyle
M D
MD
Kain
S J
SJ
Schroeder
B L
BL
Curtiss
R
R
D000595:Amino Acid Sequence; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D004269:DNA, Bacterial; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D005798:Genes, Bacterial; D006801:Humans; D008969:Molecular Sequence Data; D010958:Plasminogen / Q000302:isolation & purification / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000737:chemistry / Q000235:genetics*; D018000:Receptors, Peptide; D015183:Restriction Mapping; D016415:Sequence Alignment; D013297:Streptococcus pyogenes / Q000235:genetics* / Q000378:metabolism
D004269:DNA, Bacterial; D011956:Receptors, Cell Surface; D018000:Receptors, Peptide; C065181:plasmin receptor; D010958:Plasminogen