Hydrogen bonding between flavin and protein: a resonance Raman study

Schmidt, Jack; Coudron, P E; Thompson, A.; Watters, Kenneth L.; McFarland, James T.

doi:10.1021/bi00270a011

Cited by 46 publications

(67 citation statements)

References 23 publications

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“…The most puzzling change in the H/D-exchange experiment is the appearance of the strong band at 1290 cm 1 in D 2 O solution with excitation at 406.7, which has been related to band X occurring around 1255 cm 1 in H 2 O solution. 34 We do not observe any strong band near 1255 cm 1 in H 2 O solution in agreement with Spiro et al 31 . On the basis of relative intensities, it would be tempting to assign the band at 1231 cm 1 to band X in the H 2 O solution spectrum of GO, but this would predict a relatively large upshift of 59 cm 1 in D 2 O solution.…”

Section: Oxidized Glucose Oxidasesupporting

confidence: 93%

“…It would also require a complete reassignment of the H/D-exchange pattern, which is not supported by our data. Furthermore, not only the work by Schmidt et al 34 and Spiro et al 31 on other flavoproteins and flavins in solution but also the work by (for example) Kim and Carey 35 on RF show that the 1229 cm 1 band XI is present together with band X at 1259 cm 1 and at the other H/D-sensitive modes that can account for the observed isotope shifts. Therefore, it seems more likely that band X in GO in H 2 O solution is absent or very weak (the band at 1247 cm 1 is a possible candidate), while it can be easily observed in D 2 O solution with excitation at 406.7 nm and, to a lesser extent, at 350.7 nm, and with resonance CARS at 488.1 nm.…”

Section: Oxidized Glucose Oxidasementioning

confidence: 92%

“…Strong hydrogen-bonding to C 4 O in FCB and LMO will lower the C 4 O stretching wavenumber, while coupling of C 4 O to the N(3)-H bending motion may increase it. 26,34,35 When the coupling is lost after H/D-exchange, C 4 O shifts to its regular, lower wavenumber. In GO, hydrogen-bonding to the C(4) carbonyl is much weaker, and its stretching wavenumber is expected to be higher.…”

Section: Anionic Radical Semiquinone (Fad ž− ) In Glucose Oxidasementioning

confidence: 99%

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Resonance Raman spectra of the neutral and anionic radical semiquinones of flavin adenine dinucleotide in glucose oxidase revisited

Schelvis

Pun

Goyal

et al. 2006

J. Raman Spectrosc.

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Flavin radical semiquinones are intermediates in important physiological processes. Resonance Raman (RR) spectroscopy is an important tool to determine the interactions between these radical intermediates and their protein environment that regulate their reactivity and role in the reaction mechanisms. RR spectra of flavin radical semiquinones have been available for several flavoproteins, and those in the glucose oxidase (GO) seem significantly different from all the other available data. Since GO is often used not only as a standard for flavin-containing proteins but also in biotechnological applications, we decided to reexamine the RR spectra of the neutral and anionic radical semiquinone forms of the flavin adenine dinucleotide (FAD) cofactor in this enzyme. The new data show that the vibrational wavenumbers of the neutral and anionic radical semiquinone forms of FAD in GO are very similar to those in other flavoproteins. The discrepancies that were observed earlier seem related to contributions of the FAD in different redox and protonation states. We also obtained the first RR spectra of the oxidized FAD cofactor in GO. Analysis of the vibrations of the oxidized FAD and its anionic radical semiquinone in GO in H 2 O and D 2 O solutions indicates that the subtle differences between these spectra in GO and in other flavoproteins are related to the weak hydrogen-bonding environment of the FAD cofactor in GO.

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Section: Oxidized Glucose Oxidasesupporting

confidence: 93%

Section: Oxidized Glucose Oxidasementioning

confidence: 92%

Section: Anionic Radical Semiquinone (Fad ž− ) In Glucose Oxidasementioning

confidence: 99%

See 1 more Smart Citation

Resonance Raman spectra of the neutral and anionic radical semiquinones of flavin adenine dinucleotide in glucose oxidase revisited

Schelvis

Pun

Goyal

et al. 2006

J. Raman Spectrosc.

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“…4 shows the chemical structure and the numbering of the isoalloxazine ring, and Table I summarizes the band assignments for the oxidized and twoelectron reduced forms of yeast GR. The RR spectra of oxidized GR were previously investigated (24,25). Our study shows frequencies close to those published in the study of Schmidt et al (24).…”

Section: Rr Spectra Of the E Ox Eh 2 And Mds Species Of Yeast Grsupporting

confidence: 89%

“…The RR spectra of oxidized GR were previously investigated (24,25). Our study shows frequencies close to those published in the study of Schmidt et al (24). With reference to the RR frequencies of FAD in water, Raman diagrams were previously drawn to visualize the apoprotein effect on the high frequency modes of the isoalloxazine ring (19,26).…”

Section: Rr Spectra Of the E Ox Eh 2 And Mds Species Of Yeast Grsupporting

confidence: 77%

Electrostatic Control of the Isoalloxazine Environment in the Two-electron Reduced States of Yeast Glutathione Reductase

Picaud

Desbois

2002

Journal of Biological Chemistry

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The resonance Raman spectra of the oxidized and two-electron reduced forms of yeast glutathione reductase are reported. The spectra of the oxidized enzyme indicate a low electron density for the isoalloxazine ring. As far as the two-electron reduced species are concerned, the spectral comparison of the NADPH-reduced enzyme with the glutathione-or dithiothreitol-reduced enzyme shows significant frequency differences for the flavin bands II, III, and VII. The shift of band VII was correlated with a change in steric or electronic interaction of the hydroxyl group of a conserved Tyr with the N 10 -C 10a portion of the isoalloxazine ring. Upward shifts of bands II and III observed for the glutathione-or dithiothreitol-reduced enzyme indicate both a slight change in isoalloxazine conformation and a hydrogen bond strengthening at the N 1 and/or N 5 site(s). The formation of a mixed disulfide intermediate tends to slightly decrease the frequency of bands II, III, X, XI, and XIV. To account for the different spectral features observed for the NADPH-and glutathione-reduced species, several possibilities have been examined. In particular, we propose a hydrogen bonding modulation at the N 5 site of FAD through a variable conformation of an ammonium group of a conserved Lys residue. Changes in N 5 (flavin)-protein interaction in the two-electron reduced forms of glutathione reductase are discussed in relation to a plausible mechanism of the regulation of the enzyme activity via a variable redox potential of FAD.Glutathione reductase (GR, 1 EC 1.6.4.2, NADPH:oxidized glutathione oxidoreductase) is a pyridine nucleotide-disulfide oxidoreductase that shares many spectroscopic and structural properties with other members of this family such as lipoamide dehydrogenase, thioredoxin reductase, NADH peroxidase, and mercuric reductase (1). GR is a cytoplasmic flavoenzyme that is widely but not universally distributed in aerobic organisms. As shown in Equation 1, it catalyzes the reduction of GSSG by NADPH to form glutathione GSH.By maintaining an elevated [GSH]/[GSSG] ratio in most eukaryotic cells, GR participates in several vital functions such as the detoxification of reactive oxygen species as well as protein and DNA biosynthesis (1). Glutathione reductase has been isolated, characterized, and sequenced from different sources (1). The x-ray structures of the enzymes purified from human erythrocyte and Escherichia coli have been refined at high resolution (2-5). These structures show that GR is a homodimer. Each subunit (molecular mass ϭ 49 -59 kDa) contains one molecule of FAD and is composed of four domains, i.e. from the N-to the C-terminal end of the polypeptide chain, FAD-binding, NADPH-binding, central, and interface domains (3-5). The FAD-binding domain includes not only the FAD cofactor but also a part of the GSSG-binding site, the redox-active disulfide bridge, which is located adjacent to the si-side of the isoalloxazine ring, and a His-Glu pair considered to be the acid-base catalyst in the catalytic electron transfer (...

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Characterization of the flavin–protein interaction in L‐Lactate oxidase and old yellow enzyme by resonance inverse Raman spectroscopy

Bienstock¹,

Schopfer²,

Morris³

1987

J Raman Spectroscopy

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Resonance inverse Raman spectroscopy was used to examine the interactions between the flavin and surounding protein in L-lactate oxidase (from Mycobacterium smegmatis) and Old Yellow Enzyme (from brewer's bottom yeast). Spectra were taken of the enzymes both free and bound to various ligands. For L-lactate oxidase, the ligands consisted of substrate analogs (acetate, propionate) and inorganic anions (phosphate, sulfate and nitrate). For the inorganic anions, the expected attenuation with detuning was not observed for several bands which are associated with flavin rings I1 and 111. This effect appears to be due to a disruption of ring stacking between the uracil-pyrazine end of the flavin moiety and an aromatic amino acid. A shift in the 1232 cm-' band and changes in the bandwidths of bands in the 1450-1600 cm-' region indicate minor reorganization of the hydrogen bonding structure around the isoalloxazine on ligand binding.For Old Yellow Enzyme, the ligand was chloride. Chloride caused a slight change to the Raman spectrum, indicative of decreased hydrogen bonding to the flavin.

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Hydrogen bonding between flavin and protein: a resonance Raman study

Cited by 46 publications

References 23 publications

Resonance Raman spectra of the neutral and anionic radical semiquinones of flavin adenine dinucleotide in glucose oxidase revisited

Resonance Raman spectra of the neutral and anionic radical semiquinones of flavin adenine dinucleotide in glucose oxidase revisited

Electrostatic Control of the Isoalloxazine Environment in the Two-electron Reduced States of Yeast Glutathione Reductase

Characterization of the flavin–protein interaction in L‐Lactate oxidase and old yellow enzyme by resonance inverse Raman spectroscopy

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