Peptidyl-prolyl cis-trans isomerase from Bacillus subtilis. A prokaryotic enzyme that is highly sensitive to cyclosporin A.
Cyclophylins are members of a class of proteins with peptidyl-prolyl cis-trans isomerase activity. These enzymes bind the immunosuppressive agent, cyclosporin A (CsA), which acts as a competitive inhibitor. The peptidyl-prolyl cis-trans isomerase from Bacillus subtilis (PPIase) was purified to homogeneity in a 4-step purification procedure, which resulted in a 100-fold protein purification with a yield of 5%. Coomassie blue-stained SDS-PAGE revealed a single band of about 18 kDa. PPIase activity was determined using synthetic peptides as substrates in a 2-step reaction coupled to chymotrypsin. Treatment of Bacillus subtilis PPIase by CsA revealed an inhibition constant of Ki = 175 nM, which differs from cyclophilin of enterobacteria such as E. coli or Salmonella typhimurium and is in the range of human enzymes.
Herrler
M
M
Philips-Universität Marburg, Fachbereich Chemie/Biochemie, Germany.
Bang
H
H
Brune
K
K
Fischer
G
G
Marahiel
M A
MA
D000591:Amino Acid Isomerases / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism*; D001412:Bacillus subtilis / Q000187:drug effects / Q000201:enzymology*; D002352:Carrier Proteins / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism*; D002852:Chromatography, Ion Exchange; D016572:Cyclosporine / Q000494:pharmacology*; D004591:Electrophoresis, Polyacrylamide Gel; D007166:Immunosuppressive Agents / Q000494:pharmacology; D019696:Peptidylprolyl Isomerase
D002352:Carrier Proteins; D007166:Immunosuppressive Agents; D016572:Cyclosporine; D000591:Amino Acid Isomerases; D019696:Peptidylprolyl Isomerase