title: ESR investigations of free and immobilized glutamate dehydrogenase.
abstract: The structure of bovine liver glutamate dehydrogenase was examined with 2,2,6,6-tetramethyl-4-oxopiperidine-1-oxyl (TEMPO I) and 4-((4-(chloromercurio)benzoyl) amino)-2, 2, 6, 6-tetramethyl-1-piperidinyloxy (TEMPO II). ESR spectra from TEMPO I show that enzyme structure in the vicinity of this spin label was not distorted during immobilization to a Sepharose support. Deactivation studies with pyridoxal 5'-phosphate indicate that immobilization did not expose additional binding sites to TEMPO I. Spectra from TEMPO II reveal that immobilization profoundly altered conformational change induced by alpha-ketoglutarate and suppressed that induced by GTP and NADPH. This structural investigation provides insight into the altered kinetic properties of Sepharose-immobilized glutamate dehydrogenase and suggests a fundamental difference between monomers and allosteric oligomers in their structural response to immobilization.
authors.affiliation: Department of Chemical Engineering, California Institute of Technology, Pasadena, California 91125, USA.
authors.forename: K C
authors.initials: KC
authors.lastname: O'Connor
authors.forename: J E
authors.initials: JE
authors.lastname: Bailey