Open AccessCloning, sequencing and expression in Escherichia coli of the low‐affinity penicillin binding protein of Enterococcus faecalis
Author(s) -
Signoretto C.,
Boaretti M.,
Canepari P.
Publication year - 1994
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1994.tb07207.x
Subject(s) - enterococcus faecalis , penicillin binding proteins , biology , enterococcus hirae , gene , escherichia coli , cloning (programming) , coding region , microbiology and biotechnology , peptide sequence , biochemistry , enterococcus , antibiotics , computer science , programming language
Low‐affinity penicillin binding proteins are particular membrane proteins, in several Gram‐positive bacteria, which are involved in β‐lactam antibiotic resistance. The structural gene for the low‐affinity penicillin binding protein 5 (PBP5) of Enterococcus faecalis was cloned and sequenced. From the sequence of the 3378 bp, a 2040 bp coding region was identified. From biochemical analysis it emerges that E. faecalis PBP5 is a type II membrane protein with an uncleaved N‐terminal and is composed of 679 amino acids with a molecular weight of 74055. This protein showed 48 and 33% of identity with Enterococcus hirae PBP5 and Staphylococcus aureus PBP2a, both low‐affinity PBPs involved in β‐lactam resistance. Anti‐PBP5 antibodies cross‐reacted with a membrane protein present in other species of enterococci, but the entire gene fragment cloned hybridized only with DNAs of E. faecalis strains, thus suggesting that genes coding for low‐affinity PBPs of enterococci are not stictly homologous. In this experiment digoxigenin‐labelled E. faecalis DNA was used.