Localization of the FK506-binding protein, FKBP 13, to the lumen of the endoplasmic reticulum.
The function of the immunophilins, FKBP 12 and FKBP 13, which are binding proteins for the immunosuppressant drug FK506 and rapamycin, remains poorly defined, although it has been suggested that immunophilins and immunophilin-like proteins may play a role in protein sorting/folding and intracellular calcium ion regulation. As a first step towards understanding the function of FKBP 13, we studied its subcellular localization by immunoblotting of well-defined subcellular fractions from a canine pancreatic homogenate and immunocytochemical analysis of an overexpressed cloned cDNA for FKBP 13. Whereas FKBP 12 fractionated entirely into the cytosol, virtually all FKBP 13 was found in the rough microsomal fraction which consisted of highly purified rough endoplasmic reticulum (ER), along with several well-characterized ER markers [the immunoglobulin heavy-chain binding protein (BiP), grp 94 and ribophorin I]. Moreover, FKBP 13 co-banded with the ER markers on isopycnic sucrose gradients. By immunofluorescence, the overexpressed cDNA for FKBP 13 in Hela cells gave an ER-staining pattern highly similar to that of known ER proteins. Addition of the ligand FK506 did not appear to alter the distribution of FKBP 13. Separation of the ER luminal contents and membrane revealed FKBP 13 to be a luminal ER protein. Since the lumen of the ER is where the folding of membrane and secreted proteins occurs, as well as a major site of intracellular calcium storage, it seems possible that FKBP 13 may be involved in one of these functions.
Department of Medicine, Renal Division, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115.
S K
SK
Nigam
Y J
YJ
Jin
M J
MJ
Jin
K T
KT
Bush
B E
BE
Bierer
S J
SJ
Burakoff
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002352:Carrier Proteins / Q000032:analysis* / Q000737:chemistry / Q000235:genetics; D002500:Centrifugation, Isopycnic; D004247:DNA / Q000032:analysis / Q000235:genetics; D004285:Dogs; D004721:Endoplasmic Reticulum / Q000737:chemistry*; D005455:Fluorescent Antibody Technique; D017353:Gene Deletion; D016259:Genes, myc; D006367:HeLa Cells; D006360:Heat-Shock Proteins / Q000032:analysis* / Q000737:chemistry / Q000235:genetics; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D008969:Molecular Sequence Data; D016296:Mutagenesis; D010179:Pancreas / Q000648:ultrastructure*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D022021:Tacrolimus Binding Proteins; D014162:Transfection
D002352:Carrier Proteins; D006360:Heat-Shock Proteins; D011994:Recombinant Proteins; D004247:DNA; D022021:Tacrolimus Binding Proteins