Menaquinone as well as ubiquinone as a bound quinone crucial for catalytic activity and intramolecular electron transfer in Escherichia coli membrane-bound glucose dehydrogenase.
Escherichia coli membrane-bound glucose dehydrogenase (mGDH), which is one of quinoproteins containing pyrroloquinoline quinone (PQQ) as a coenzyme, is a good model for elucidating the function of bound quinone inside primary dehydrogenases in respiratory chains. Enzymatic analysis of purified mGDH from cells defective in synthesis of ubiquinone (UQ) and/or menaquinone (MQ) revealed that Q-free mGDH has very low levels of activity of glucose dehydrogenase and UQ2 reductase compared with those of UQ-bearing mGDH, and both activities were significantly increased by reconstitution with UQ1. On the other hand, MQ-bearing mGDH retains both catalytic abilities at the same levels as those of UQ-bearing mGDH. A radiolytically generated hydrated electron reacted with the bound MQ to form a semiquinone anion radical with an absorption maximum at 400 nm. Subsequently, decay of the absorbance at 400 nm was accompanied by an increase in the absorbance at 380 nm with a first order rate constant of 5.7 x 10(3) s(-1). This indicated that an intramolecular electron transfer from the bound MQ to the PQQ occurred. EPR analysis revealed that characteristics of the semiquinone radical of bound MQ are similar to those of the semiquinone radical of bound UQ and indicated an electron flow from PQQ to MQ as in the case of UQ. Taken together, the results suggest that MQ is incorporated into the same pocket as that for UQ to perform a function almost equivalent to that of UQ and that bound quinone is involved at least partially in the catalytic reaction and primarily in the intramolecular electron transfer of mGDH.
Mustafa
Golam
G
Department of Biological Chemistry, Faculty of Agriculture, Yamaguchi University, Yamaguchi 753-8515, Japan.
Migita
Catharina T
CT
Ishikawa
Yoshinori
Y
Kobayashi
Kazuo
K
Tagawa
Seiichi
S
Yamada
Mamoru
M
D000838:Anions; D002384:Catalysis; D004578:Electron Spin Resonance Spectroscopy; D004579:Electron Transport / Q000187:drug effects; D004583:Electrons; D004926:Escherichia coli / Q000378:metabolism*; D005609:Free Radicals; D042843:Glucose 1-Dehydrogenase / Q000378:metabolism*; D007700:Kinetics; D008280:Magnetics; D011809:Quinones / Q000737:chemistry; D013997:Time Factors; D014451:Ubiquinone / Q000378:metabolism*; D024482:Vitamin K 2 / Q000378:metabolism*; D014815:Vitamins / Q000378:metabolism*
D000838:Anions; D005609:Free Radicals; D011809:Quinones; D014815:Vitamins; D024482:Vitamin K 2; D014451:Ubiquinone; D042843:Glucose 1-Dehydrogenase