Differential gene expression and extracellular secretion of the collagenolytic enzymes by the pathogen Vibrio parahaemolyticus.
Vibrio parahaemolyticus, a causative agent of wound infections as well as food poisoning, harbors two collagenase genes: vppC and prtV. When cultivated at 26 degrees C in gelatin broth supplemented with 3.0% NaCl, significant collagenolytic activity was detected in the culture supernatant at the early stationary phase. Native polyacrylamide gel electrophoresis analysis revealed a 90-kDa protein, and N-terminal amino acid sequencing showed that this protein was VppC, generated through truncation of 72 N-terminal amino acid residues. Additionally, significant expression of only vppC was observed by reverse transcriptase PCR. By contrast, a vppC-negative mutant constructed through single crossover homologous recombination secreted a 50-kDa-collagenolytic enzyme; however, this enzyme was a serine protease that was reported previously. These results suggest that VppC is a primary extracellular collagenase produced by V. parahaemolyticus.
Miyoshi
Shin-ichi
S
Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Tushima-Naka, Okayama, Japan. miyoshi@pharm.okayama-u.ac.jp
Nitanda
Yuko
Y
Fujii
Kaori
K
Kawahara
Kiyomi
K
Li
Tao
T
Maehara
Yoko
Y
Ramamurthy
Thandavarayan
T
Takeda
Yoshifumi
Y
Shinoda
Sumio
S
D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D017364:Collagenases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D003470:Culture Media / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D017353:Gene Deletion; D020869:Gene Expression Profiling; D008970:Molecular Weight; D012329:RNA, Bacterial / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D020539:Sequence Analysis, Protein; D012965:Sodium Chloride; D013696:Temperature; D014736:Vibrio parahaemolyticus / Q000201:enzymology*
D001426:Bacterial Proteins; D003470:Culture Media; D012329:RNA, Bacterial; D012333:RNA, Messenger; D012965:Sodium Chloride; D017364:Collagenases