title: Study on the interactions between protein disulfide isomerase and target proteins, using immobilization on solid support.
abstract: Interaction between protein disulfide isomerase, possessing not only isomerase but also chaperone-like activity, and olygomeric enzyme, GAPDH, has been studied using technique of immobilization on insoluble support. PDI dimers bound to CNBr-activated Sepharose were shown to possess high TPOR activity as well as the ability to reactivate lysozyme. Immobilized PDI was not found to interact neither with soluble tetrameric GAPDH, nor with soluble denatured GAPDH. However, soluble PDI binds effectively to immobilized GAPDH monomers; Kd was found to be 3.7 x 10(-6) M, stoichiometry 0.824 mole PDI monomers per mole GAPDH monomers. Immobilized GAPDH tetramers do not interact with PDI. These observations are also confirmed by the data on electrophoresis of proteins bound to immobilized GAPDH monomers and tetramers. The ability of PDI to interact with denatured protein form, but not with the native one, is considered to be evidence of chaperone-like activity of the enzyme.
authors.affiliation: A.N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, Russia. muric@bac.genebee.msu.su
authors.forename: V I
authors.initials: VI
authors.lastname: Muronetz
authors.forename: N X
authors.initials: NX
authors.lastname: Zhang
authors.forename: I G
authors.initials: IG
authors.lastname: Bulatnikov
authors.forename: C C
authors.initials: CC
authors.lastname: Wang
mesh_terms: D004220:Disulfides; D004800:Enzymes, Immobilized; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000737:chemistry / Q000378:metabolism*; D018832:Molecular Chaperones; D009113:Muramidase / Q000378:metabolism; D011485:Protein Binding; D011489:Protein Denaturation; D019704:Protein Disulfide-Isomerases / Q000378:metabolism*; D012995:Solubility; D013329:Structure-Activity Relationship
chemical_list: D004220:Disulfides; D004800:Enzymes, Immobilized; D018832:Molecular Chaperones; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D009113:Muramidase; D019704:Protein Disulfide-Isomerases