Expression cloning and biochemical characterizations of recombinant cyclophilin proteins from Schistosoma mansoni.
Recombinant Schistosoma mansoni cyclophilin proteins of the A and the B subtypes (SmCYP A and B) were expressed in bacterial cells as histidine- and maltose-binding fusion proteins and also as nonfused proteins. In addition, S. mansoni CYPs were produced in Sf9 insect cells in their natural forms. Purified recombinant SmCYP B was found to possess a peptidyl-prolyl cis-trans isomerase (PPIase) activity, with a kcat/Km value of 8.2 x 10(5) M-1 s-1. The SmCYP B isoform is approximately two to three times more active than SmCYP A. SmCYP B-specific RNA appears to be more abundant in adult schistosomes than SmCYP A RNA in Northern blots. These results support the conclusion that SmCYP B represents the major schistosomal CYP. The PPIase-associated activity of both CYPs was inhibitable by the immunosuppressive drug cyclosporin A (CsA). We attempt to explain differences in PPIase activities and in CsA inhibition by examining models of the two CYPs complexed to CsA.
Bugli
F
F
Istituto di Biologia Cellulare, Consiglio Nazionale delle Ricerche, 43 Viale Marx, Rome, 00137, Italy.
Khattab
A
A
Vigneti
E
E
Butler
R
R
Cioli
D
D
Klinkert
M Q
MQ
D000818:Animals; D001665:Binding Sites; D015153:Blotting, Western; D002460:Cell Line; D003001:Cloning, Molecular; D016572:Cyclosporine / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006639:Histidine / Q000737:chemistry; D007106:Immune Sera / Q000276:immunology; D008320:Maltose / Q000737:chemistry; D008958:Models, Molecular; D019696:Peptidylprolyl Isomerase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D011487:Protein Conformation; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D012550:Schistosoma mansoni / Q000201:enzymology*
D004791:Enzyme Inhibitors; D007106:Immune Sera; D011993:Recombinant Fusion Proteins; D006639:Histidine; D008320:Maltose; D016572:Cyclosporine; D019696:Peptidylprolyl Isomerase