Hormone binding by protein disulfide isomerase, a high capacity hormone reservoir of the endoplasmic reticulum.
Protein disulfide isomerase (PDI) is a folding assistant of the eukaryotic endoplasmic reticulum, but it also binds the hormones, estradiol, and 3,3',5-triiodo-l-thyronine (T(3)). Hormone binding could be at discrete hormone binding sites, or it could be a nonphysiological consequence of binding site(s) that are involved in the interaction PDI with its peptide and protein substrates. Equilibrium dialysis, fluorescent hydrophobic probe binding (4,4'-dianilino-1,1'-binaphthyl-5,5'-disulfonic acid (bis-ANS)), competition binding, and enzyme activity assays reveal that the hormone binding sites are distinct from the peptide/protein binding sites. PDI has one estradiol binding site with modest affinity (2.1 +/- 0.5 microm). There are two binding sites with comparable affinity for T(3) (4.3 +/- 1.4 microm). One of these overlaps the estradiol site, whereas the other binds the hydrophobic probe, bis-ANS. Neither estradiol nor T(3) inhibit the catalytic or chaperone activity of PDI. Although the affinity of PDI for the hormones estradiol and T(3) is modest, the high local concentration of PDI in the endoplasmic reticulum (>200 microm) would drive hormone binding and result in the association of a substantial fraction (>90%) of the hormones in the cell with PDI. High capacity, low affinity hormone sites may function to buffer hormone concentration in the cell and allow tight, specific binding to the true receptor while preserving a reasonable number of hormone molecules in the very small volume of the cellular environment.
Primm
T P
TP
Verna and Marrs McLean Department of Biochemistry, Baylor College of Medicine, Houston, Texas 77030, USA.
Gilbert
H F
HF
D000817:Anilino Naphthalenesulfonates / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D001414:Bacitracin / Q000494:pharmacology; D001667:Binding, Competitive; D002384:Catalysis / Q000187:drug effects; D004721:Endoplasmic Reticulum / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D004958:Estradiol / Q000737:chemistry / Q000378:metabolism*; D008024:Ligands; D018832:Molecular Chaperones / Q000378:metabolism; D011485:Protein Binding; D019704:Protein Disulfide-Isomerases / Q000378:metabolism*; D017510:Protein Folding; D051381:Rats; D013050:Spectrometry, Fluorescence; D014284:Triiodothyronine / Q000737:chemistry / Q000378:metabolism*
D000817:Anilino Naphthalenesulfonates; D008024:Ligands; D018832:Molecular Chaperones; D014284:Triiodothyronine; D001414:Bacitracin; D004958:Estradiol; C035763:5,5'-bis(8-(phenylamino)-1-naphthalenesulfonate); D019704:Protein Disulfide-Isomerases