title: Relationship between the conformation of glutamate dehydrogenase, the state of association of its subunit, and catalytic function.
abstract: The fluorescence and phosphorescence properties of the tryptophan residues in glutamate dehydrogenase were utilized to probe the conformation of the macromolecule at various states of aggregation of its subunits (hexamer, trimer, and monomer) in guanidine hydrochloride. According to the phosphorescence lifetime no gross alteration in the conformation of the protein follows from complete dissociation of the hexamer into native monomer, implying that the native fold is stabilized exclusively by intrasubunit bonding. Although modest concentrations of denaturant induce a change in configuration in the enzyme, a comparison with the macromolecule cross-linked into the hexameric form by glutaraldehyde confirms that this alteration in structure is not the result of subunit dissociation. Inhibition of catalysis by the denaturant is found to be considerably smaller than anticipated from the extent of hexamer dissociation. Furthermore, this inhibition is in no way prevented by cross-linking the enzyme in its hexameric form. This finding together with the ability of the trimer to bind the coenzyme and to undergo the characteristic structural changes induced by the effectors ADP and GTP suggests that, contrary to what is generally believed, the smallest functional unit of glutamate dehydrogenase is not the hexameric form.
authors.affiliation: Istituto di Biofisica, CNR, Pisa, Italy.
authors.forename: G B
authors.initials: GB
authors.lastname: Strambini
authors.forename: P
authors.initials: P
authors.lastname: Cioni
authors.forename: A
authors.initials: A
authors.lastname: Puntoni
mesh_terms: D000818:Animals; D002417:Cattle; D005969:Glutamate Dehydrogenase / Q000378:metabolism*; D019791:Guanidine; D006146:Guanidines / Q000494:pharmacology; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008163:Luminescent Measurements; D046911:Macromolecular Substances; D009994:Osmolar Concentration; D011487:Protein Conformation; D011789:Quantum Theory; D013050:Spectrometry, Fluorescence
chemical_list: D006146:Guanidines; D046911:Macromolecular Substances; D005969:Glutamate Dehydrogenase; D019791:Guanidine