Characterization of cyclophilin 40: Highly conserved protein that directly associates with Hsp90

Abstract

Cyclophilin 40 (CyP40) is a recently identified member of the cyclophilin family that may be a component of unactivated steroid receptor complexes. It consists of an N-half portion that is highly homologous to cyclophilin A and has peptidyl prolyl isomerase (PPIase) activity, and a C-half portion that resembles the C- terminal portion of FKBP52 (FK506 binding protein 52), another component of unactivated steroid receptor complexes. To better understand the structure and functional characteristics of this new class of cyclophilin, we have raised monoclonal antibodies against the C-half portion of human CyP40. Immunostaining with the antibodies showed its preferential localization in cytoplasm. One antibody cross-reacted with a 45kDa protein in yeast, suggesting high conservation throughout evolution. A CyP40-associated protein was isolated from rabbit reticulocyte lysate by means of an affinity resin, and was identified as hsp90. The C-half portion of CyP40 was necessary and sufficient for the interaction.