Isolation and characterization of a 40-kDa cyclophilin-related protein.
Major and minor isoforms of cyclophilin (CyP-18), a 17.8-kDa protein with peptidyl-prolyl cis-trans isomerase activity, comprise the primary intracellular binding proteins for cyclosporin A. Additional CyP-like proteins with approximate molecular masses of 22 (CyP-22) and 40 kDa (CyP-40) have been recovered from the soluble fraction of calf brain along with CyP-18 by adsorption onto a cyclosporin A affinity column and elution with cyclosporin A. Based on a limited number of peptide sequences from CyP-22, it appears that we have isolated from tissue CyPB, a protein whose sequence was deduced previously from cloned cDNA. The 40-kDa protein was separated from CyP-18 and CyP-22 on a molecular sieving column. Isoelectric focusing of CyP-40 yielded two bands at pI 5.3 and 5.5, in contrast to the basic pI values of CyP-18. Some tryptic peptides from CyP-40 were found to be highly homologous but not identical to bovine CyP-18; others were not significantly homologous to CyP-18 or any other protein in the data base. Unlike the major and minor isoforms of Cyp-18, monospecific polyclonal anti-CyP-18 antibodies did not cross-react with CyP-22 and CyP-40. Likewise, anti-CyP-40 serum minimally cross-reacts with CyP-18 and CyP-22. Cyp-40 possesses peptidyl-prolyl cis-trans isomerase activity which is less sensitive to inhibition by cyclosporin A (IC50 = 300 nM) than is CyP-18 (IC50 = 20 nM).
Kieffer
L J
LJ
Yale University School of Medicine, New Haven, Connecticut 06510.
Thalhammer
T
T
Handschumacher
R E
RE
D000591:Amino Acid Isomerases / Q000302:isolation & purification* / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism*; D002352:Carrier Proteins / Q000302:isolation & purification* / Q000378:metabolism; D002417:Cattle; D002645:Chickens; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D003524:Cyclosporins / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D007106:Immune Sera; D007525:Isoelectric Focusing; D008969:Molecular Sequence Data; D008970:Molecular Weight; D019696:Peptidylprolyl Isomerase; D012689:Sequence Homology, Nucleic Acid; D013950:Thymus Gland / Q000378:metabolism
D002352:Carrier Proteins; D003524:Cyclosporins; D007106:Immune Sera; D000591:Amino Acid Isomerases; D019696:Peptidylprolyl Isomerase