title: Interaction of BiP with substance P and nucleotides.
abstract: A rapid and simple spin column assay has been used to study interactions of BiP with substance P (SP) and ATP. At 4 degrees C, the binding of SP to BiP requires ATP and a stable SP-BiP.ATP complex is formed. Nonhydrolyzable ATP analogues or ADP cannot replace ATP. Although ATP converts BiP dimers to monomers, the requirement for ATP for SP binding is not solely due to BiP dissociation, because purified BiP monomers also require ATP for peptide binding. At 37 degrees C, there is rapid binding of SP to BiP even in the absence of ATP and, in fact, ATP at concentrations above 5 microM causes release of SP from BiP. At this higher temperature, there is also rapid hydrolysis of ATP bound to BiP. These results extend our previous results (Brot et al., 1994) that indicated the formation, at low ATP concentrations, of a labile SP.BiP.ATP complex that, after ATP hydrolysis, resulted in a stable SP.BiP.ADP complex.
authors.affiliation: Roche Institute of Molecular Biology, Roche Research Center, Nutley, NJ 07110, USA.
authors.forename: H
authors.initials: H
authors.lastname: Weissbach
authors.forename: B
authors.initials: B
authors.lastname: Redfield
authors.forename: N
authors.initials: N
authors.lastname: Qiu
authors.forename: G
authors.initials: G
authors.lastname: Chen
authors.forename: A
authors.initials: A
authors.lastname: Carlino
authors.forename: V
authors.initials: V
authors.lastname: Vidal
authors.forename: O
authors.initials: O
authors.lastname: Tsolas
authors.forename: N
authors.initials: N
authors.lastname: Brot
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism*; D002352:Carrier Proteins / Q000737:chemistry / Q000378:metabolism*; D002850:Chromatography, Gel / Q000379:methods; D003911:Dextrans; D000091342:Endoplasmic Reticulum Chaperone BiP; D006360:Heat-Shock Proteins / Q000737:chemistry / Q000378:metabolism*; D006868:Hydrolysis; D008099:Liver / Q000737:chemistry; D018832:Molecular Chaperones / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000378:metabolism; D013373:Substance P / Q000378:metabolism*; D013696:Temperature
chemical_list: D002352:Carrier Proteins; D003911:Dextrans; D000091342:Endoplasmic Reticulum Chaperone BiP; D006360:Heat-Shock Proteins; D018832:Molecular Chaperones; D011994:Recombinant Proteins; D013373:Substance P; D000255:Adenosine Triphosphate; C025614:sephadex