title: Purification and characterization of an oxygen-stable form of dinitrogenase reductase-activating glycohydrolase from Rhodospirillum rubrum.
abstract: Dinitrogenase reductase-activating glycohydrolase (DRAG) is responsible for removing the ADP-ribose moiety from post-translationally inactivated nitrogenase of Rhodospirillum rubrum. Using DRAG purified from an overexpressing strain (UR276), further properties of this enzyme were studied, including its u.v.-visible and fluorescence spectra and its stability in air. DRAG appears to require no covalently bound inorganic cofactors for its activity or regulation. Previously, purified DRAG was found to be rapidly inactivated in air. The air-catalysed lability originated with the presence of sodium dithionite and Mn2+ throughout the purification of the enzyme. This lability can be mimicked using H2O2, which is known to oxidatively inactivate proteins containing bivalent metals. Implications for the regulation of nitrogenase are discussed with respect to the lack of sensitivity to air of the regulatory enzyme, DRAG.
authors.affiliation: Department of Biochemistry, University of Wisconsin-Madison 53706.
authors.forename: G M
authors.initials: GM
authors.lastname: Nielsen
authors.forename: Y
authors.initials: Y
authors.lastname: Bao
authors.forename: G P
authors.initials: GP
authors.lastname: Roberts
authors.forename: P W
authors.initials: PW
authors.lastname: Ludden
mesh_terms: D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D006026:Glycoside Hydrolases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D006861:Hydrogen Peroxide / Q000494:pharmacology; D009699:N-Glycosyl Hydrolases; D009586:Nitrogen Fixation; D010100:Oxygen / Q000494:pharmacology; D012247:Rhodospirillum rubrum / Q000201:enzymology*; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet
chemical_list: D006861:Hydrogen Peroxide; D006026:Glycoside Hydrolases; D009699:N-Glycosyl Hydrolases; C036091:ADP-ribosyl-(dinitrogen reductase) hydrolase; D010100:Oxygen