Isolation and characterization of a 17-kDa FKBP-type peptidyl-prolyl cis/trans isomerase from Vibrio anguillarum.
Peptidyl-prolyl cis/trans isomerase (PPIase) catalyzes the isomerization of peptide bonds to achieve conformational changes in native folded proteins. An FKBP-type PPIase with an approximate molecular weight of 17kDa was isolated from Vibrio anguillarum O1 and named VaFKBP17. To investigate its biochemical properties, the ppi gene from V. anguillarum O1 was isolated and overexpressed in Escherichia coli. A protease-coupled assay for isomerization activity, using Succinyl-Ala-Phe-Pro-Phe-p nitroanilide as substrate, indicated that the activity of VaFKBP17 was highest at low temperature (5°C) and alkaline conditions (pH 10). The immunosuppressant FK506 inhibited the isomerization activity of VaFKBP17. The chaperone activity of VaFKBP17 was assessed using a citrate synthase thermal aggregation activity assay. To evaluate its ability to catalyze protein refolding, the effect of VaFKBP17 on inclusion bodies was investigated during a dilution process. In this assay, VaFKBP17 was able to assist protein refolding. These results provide evidence that VaFKBP17 possesses chaperone-like activity. The structural homology of VaFKBP17 relative to other known bacterial FKBPs was also examined.
Department of Biotechnology, Pukyong National University, Busan 608-737, Republic of Korea.
Geon-A
GA
Jo
Department of Biotechnology, Pukyong National University, Busan 608-737, Republic of Korea.
Jong Min
JM
Lee
Department of Biotechnology, Pukyong National University, Busan 608-737, Republic of Korea.
Gyuyou
G
No
Department of Biotechnology, Pukyong National University, Busan 608-737, Republic of Korea.
Dong Seop
DS
Kang
Department of Biotechnology, Pukyong National University, Busan 608-737, Republic of Korea.
So-Hyun
SH
Kim
Department of Oral Biochemistry, Dental Science Research Institute, Medical Research Center for Biomineralization Disorders, School of Dentistry, Chonnam National University, Gwangju, Republic of Korea.
Sun-Hee
SH
Ahn
Department of Biotechnology, Pukyong National University, Busan 608-737, Republic of Korea. Electronic address: iskong@pknu.ac.kr.
In-Soo
IS
Kong
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006639:Histidine / Q000737:chemistry / Q000235:genetics; D006863:Hydrogen-Ion Concentration; D002479:Inclusion Bodies / Q000737:chemistry / Q000201:enzymology; D008958:Models, Molecular; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009842:Oligopeptides / Q000737:chemistry / Q000235:genetics; D019696:Peptidylprolyl Isomerase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism*; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D016415:Sequence Alignment; D040681:Structural Homology, Protein; D013379:Substrate Specificity; D016559:Tacrolimus / Q000737:chemistry; D013696:Temperature; D014733:Vibrio / Q000737:chemistry* / Q000201:enzymology
D001426:Bacterial Proteins; C471213:His-His-His-His-His-His; D009842:Oligopeptides; D011993:Recombinant Fusion Proteins; D006639:Histidine; D019696:Peptidylprolyl Isomerase; D016559:Tacrolimus
FK506-binding protein; Peptidyl-prolyl cis/trans isomerase; Single domain; Vibrio anguillarum