Different regions of the immunophilin FKBP52 determine its association with the glucocorticoid receptor, hsp90, and cytoplasmic dynein.
FKBP52 is a high molecular mass immunophilin possessing peptidylprolyl isomerase (PPIase) activity that is inhibited by the immunosuppressant drug FK506. FKBP52 is a component of steroid receptor.hsp90 heterocomplexes, and it binds to hsp90 via a region containing three tetratricopeptide repeats (TPRs). Here we demonstrate by cross-linking of the purified proteins that there is one binding site for FKBP52/dimer of hsp90. This accounts for the common heterotetrameric structure of native receptor heterocomplexes being 1 molecule of receptor, 2 molecules of hsp90, and 1 molecule of a TPR domain protein. Immunoadsorption of FKBP52 from reticulocyte lysate also yields co-immunoadsorption of cytoplasmic dynein, and we show that co-immunoadsorption of dynein is competed by a fragment of FKBP52 containing its PPIase domain, but not by a TPR domain fragment that blocks FKBP52 binding to hsp90. Using purified proteins, we also show that FKBP52 binds directly to the hsp90-free glucocorticoid receptor. Because neither the PPIase fragment nor the TPR fragment affects the binding of FKBP52 to the glucocorticoid receptor under conditions in which they block FKBP52 binding to dynein or hsp90, respectively, different regions of FKBP52 must determine its association with these three proteins.
Silverstein
A M
AM
Department of Pharmacology, University of Michigan Medical School, Ann Arbor, Michigan 48109, USA.
Galigniana
M D
MD
Kanelakis
K C
KC
Radanyi
C
C
Renoir
J M
JM
Pratt
W B
WB
D000818:Animals; D001665:Binding Sites; D002478:Cells, Cultured; D004398:Dyneins / Q000378:metabolism*; D018841:HSP90 Heat-Shock Proteins / Q000378:metabolism*; D020104:Immunophilins / Q000737:chemistry / Q000378:metabolism*; D011965:Receptors, Glucocorticoid / Q000378:metabolism*; D018411:Spodoptera; D022021:Tacrolimus Binding Proteins
D018841:HSP90 Heat-Shock Proteins; D011965:Receptors, Glucocorticoid; D004398:Dyneins; D022021:Tacrolimus Binding Proteins; D020104:Immunophilins