Confirmation of Vpr as a fibrinolytic enzyme present in extracellular proteins of Bacillus subtilis.
We have previously reported a proteomic approach to detect fibrinolytic enzymes from the secreted proteins of Bacillus subtilis 168 and identified two extracellular fibrinolytic enzymes of Bacillus, namely, Vpr and WprA. In this study, to confirm the fibrinolytic activity of Vpr, we cloned the vpr gene and expressed it in Escherichia coli, where it is predominantly localized to inclusion bodies. After affinity purification and desalting steps, the expressed Vpr is auto-processed to an active form. Interestingly, after the desalting step, several additional bands with fibrinolytic activity were detected in zymography gel along with a mature form (68 kDa) of Vpr. MALDI-TOF analyses of these bands revealed that Vpr could exist in multiple forms.
Kho
Chang Won
CW
Systemic Proteomics Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Yuseong, P.O. Box 115, Daejeon 305-600, South Korea.
Park
Sung Goo
SG
Cho
Sayeon
S
Lee
Do Hee
DH
Myung
Pyung Keun
PK
Park
Byoung Chul
BC
D001412:Bacillus subtilis / Q000201:enzymology*; D001426:Bacterial Proteins / Q000302:isolation & purification / Q000378:metabolism*; D004229:Dithiothreitol / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D012697:Serine Endopeptidases / Q000302:isolation & purification / Q000378:metabolism*; D014508:Urea / Q000378:metabolism
D001426:Bacterial Proteins; D014508:Urea; D012697:Serine Endopeptidases; C501685:Vpr protein, Bacillus subtilis; D004229:Dithiothreitol