Electron transfer from the aminosemiquinone reaction intermediate of methylamine dehydrogenase to amicyanin.
The tryptophan tryptophylquinone (TTQ) cofactor of methylamine dehydrogenase (MADH) is covalently modified by substrate-derived nitrogen during its two-electron reduction by methylamine to form an aminoquinol (N-quinol). An N-semiquinone, which retains the substrate-derived N, is the intermediate during the two sequential one-electron oxidations of N-quinol MADH by its physiologic electron acceptor, amicyanin. Electron transfer (ET) from N-quinol MADH to amicyanin is gated by the deprotonation of the substrate-derived amino group on TTQ in the enzyme active site, whereas ET reactions from dithionite-reduced quinol and semiquinone forms of MADH are rate-limited by the ET event. The ET reaction from the N-semiquinone intermediate is shown not to be gated, but rate-limited by the ET step. Marcus analysis of the reaction reveals that the ET reaction from the N-semiquinone MADH to amicyanin exhibits the same reorganizational energy and electronic coupling as do the ET reactions of the dithionite-reduced O-quinol and O-semiquinone forms. The rates of the ET reactions of these three different redox forms of MADH exhibit a DeltaG degrees dependence which is predicted by Marcus theory. The ET reaction of the N-semiquinone is relatively insensitive to pH and salt, and does not exhibit a primary kinetic solvent isotope effect over the range of pH and cation concentrations studied. These properties are similar to those of the ET reaction of quinol MADH and different from those of the gated reaction of N-quinol MADH, whose rate varies considerably with pH and concentrations of specific monovalent cations. Thus, the covalent incorporation of substrate-derived N into TTQ is not alone sufficient to cause gating of ET. It affects the rate and DeltaG degrees for the ET reaction from the TTQ semiquinone by altering its redox potential, but it does not alter the reorganizational energy and electronic coupling associated with ET from TTQ to amicyanin.
Department of Biochemistry, The University of Mississippi Medical Center, Jackson, Mississippi 39216-4505, USA. rbishop@college.emory.edu
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D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D004579:Electron Transport; D006863:Hydrogen-Ion Concentration; D045563:Indolequinones; D007700:Kinetics; D010084:Oxidation-Reduction; D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000737:chemistry / Q000378:metabolism*; D010231:Paracoccus denitrificans / Q000201:enzymology; D011189:Potassium Chloride; D011522:Protons; D011809:Quinones / Q000737:chemistry / Q000378:metabolism*; D012997:Solvents; D014364:Tryptophan / Q000031:analogs & derivatives / Q000378:metabolism
D001426:Bacterial Proteins; D045563:Indolequinones; D011522:Protons; D011809:Quinones; D012997:Solvents; C017079:mauC protein, Methylobacterium extorquens; C070569:tryptophan tryptophylquinone; D011189:Potassium Chloride; D014364:Tryptophan; C021567:methylamine dehydrogenase; D000587:Oxidoreductases Acting on CH-NH Group Donors