Effects of environment on the folding of nicotinamide-adenine dinucleotides in aqueous solutions

McDonald, George G.; Brown, Barry L.; Hollis, Donald P.; Walter, Charles

doi:10.1021/bi00760a029

Cited by 44 publications

(56 citation statements)

References 15 publications

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“…The poor efficiency of NADH relative to NMNH in the formation of IDOFe 2ϩ -O 2 may be related to the equilibrium of unfolded and folded (formed by stacking of the adenine and nicotinamide groups) conformations that NADH exhibits in solution (32,90,91). Presumably, the folded form interacts less efficiently with the active site of IDO and/or inhibits reaction of the nicotinamide with dioxygen.…”

Section: Discussionmentioning

confidence: 99%

NADH Oxidase Activity of Indoleamine 2,3-Dioxygenase

Rosell

Kuo²,

Mauk

2011

Journal of Biological Chemistry

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The heme enzyme indoleamine 2,3-dioxygenase (IDO) was found to oxidize NADH under aerobic conditions in the absence of other enzymes or reactants. This reaction led to the formation of the dioxygen adduct of IDO and supported the oxidation of Trp to N-formylkynurenine. Formation of the dioxygen adduct and oxidation of Trp were accelerated by the addition of small amounts of hydrogen peroxide, and both processes were inhibited in the presence of either superoxide dismutase or catalase. Anaerobic reaction of IDO with NADH proceeded only in the presence of a mediator (e.g. methylene blue) and resulted in formation of the ferrous form of the enzyme. We propose that trace amounts of peroxide previously proposed to occur in NADH solutions as well as solid NADH activate IDO and lead to aerobic formation of superoxide and the reactive dioxygen adduct of the enzyme.

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Section: Discussionmentioning

confidence: 99%

NADH Oxidase Activity of Indoleamine 2,3-Dioxygenase

Rosell

Kuo²,

Mauk

2011

Journal of Biological Chemistry

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“…It is therefore predictable that positioning of the eadenosine of any fluorescent coenzyme moiety (e.q., EATP, EADP) in close proximity to a tryptophan in a protein will result in complete fluorescence quenching of the former. Accumulated evidence on the conformation of nicotinamide adenine dinucleotide, NAD+, and the reduced form, NADH, in neutral aqueous solution is consistent with a population of open forms, in which the ribose-diphosphate-ribose backbone is extended, and closed forms, in which the heterocyclic rings are "stacked" in close proximity (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14). Energy transfer from adenine to dihydronicotinamide in NADH in very dilute solution is compatible only with a structure in which the two heterocyclic rings are in close proximity (15,16 (25,26) with chloroacetaldehyde, thus providing a model in which indole is used as a neutral substitute for tryptophan.…”

mentioning

confidence: 91%

Dynamic and static quenching of 1,N6-ethenoadenine fluorescence in nicotinamide 1,N6-ethenoadenine dinucleotide and in 1,N6-etheno-9-(3-(indol-3-yl) propyl) adenine.

Gruber¹,

Leonard²

1975

Proc. Natl. Acad. Sci. U.S.A.

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For nicotinamide 1,N6-ethenoadenine dinucleotide (ENAD+), the fluorescent analog of NAD+, in neutral aqueous solution the quantum yield has been determined to be 0.028 and the fluorescent lifetime, 2.1 nsec. Simultaneous determination of quantum yields and lifetimes of eNAD+ and of the "half molecule" EAMP allows the calculation of the percentage of stacked and open conformations of the dinucleotide. At 250 in neutral aqueous solution there is 45 I 5% of stacked forms. The value of the fluorescent lifetime observed for ENAD+ is sensitive to fluorescent impurities, especially those containing the E-adenosine moiety, and a purification procedure using high performance liquid chromatography was devised to obtain fluorescently homogeneous preparations.In order to study the effect on e-adenosine fluorescence caused by the possible close proximity of a tryptophan in a polypeptide chain or protein, we have prepared 1,N6-etheno- These studies relate to intramolecular stacking interactions within the ENAD+ molecule and set a probable upper limit for the interactions within the NAD+ molecule itself. It is also important to delineate possible interactions between the adenosine moiety of various coenzymes and the individual amino acids, e.g., tryptophan, of various proteins. To do so on a simplified level we synthesized 1,N6-etheno-9-[3- (indol-3-yl)propyl]adenine (EAde9-Cs-Ind3) by reaction of 9-[3-(indol-3-yl)propyl]adenine (Ade9-C3-Ind3) (25, 26) with chloroacetaldehyde, thus providing a model in which indole is used as a neutral substitute for tryptophan. From simultaneous measurement of the fluorescence lifetime and the quantum yield of EAde9-C3-Ind3 relative to those of the "half molecules" 1,N6-etheno-9-propyladenine (EAde9-C3) and 3-propylindole (Ind3-Cs), we are able to predict the fluorescence quenching effect of bringing the E-adenine moiety into close proximity with the indole of tryptophan. (27.6 MPa). The eluate was monitored by absorbance at 254 nm, recorded on a Hewlett-Packard recorder modified as described previously (27). As a criterion of purity we used identical lifetimes determined by both phase shift and modulation (28,29). This technique is very sensitive to the presence of more than a single fluorescent decay, and the presence of EAMP in our preparation would be expected to 3966

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“…Upfield shifts due to concentration-dependent stacking interaction can be neglected at these coenzyme concentrations (13). Similar studies on the binding of DPN+ to several other dehydrogenases were also performed.…”

Section: Methodsmentioning

confidence: 92%

“…Previous studies have indicated that the diphosphopyridine coenzymes exist in fast equilibrium between the open and the folded conformations in neutral aqueous solutions (13)(14)(15). In the folded conformation, both adenine and pyridine moieties of the coenzymes are stacked vertically.…”

Section: Discussionmentioning

confidence: 99%

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Conformations of Diphosphopyridine Coenzymes upon Binding to Dehydrogenases

Lee¹,

Eichner²,

Kaplan³

1973

Proc. Natl. Acad. Sci. U.S.A.

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The binding of oxidized as well as reduced coenzyme to some dehydrogenases has been studied under different concentration ratios and temperatures by nuclear magnetic resonance spectroscopy. A significant difference in the spectral behavior between DPN+ and DPNH upon binding is interpreted in terms of fast and slow on-off rates relative to the nuclear magnetic resonance time scale in the binding of these two coenzymes. Significant downfield shifts of DPN+ were observed upon binding, comparable in magnitude to those expected upon opening (destacking) of the coenzymes in the case of chicken-muscle and lobster-tail lactate dehydrogenase (EC 1.1.1.27) and yeast alcohol dehydrogenase (EC 1.1.1.1). A preliminary survey of several other dehydrogenases is consistent with these findings. In the case of 3-phosphoglyceraldehyde dehydrogenase, there is a possibility that the coenzyme exists in the folded form.Nuclear magnetic resonance spectroscopy (NMR) is now extensively applied to probe the active sites of enzymes in solution (1, 2). Most of these studies have been concerned with reversible enzyme-inhibitor or substrate interactions (3-8).Information about the active sites is often inferred from the effects that the enzyme has on the chemical shifts (9, 10), and line-widths (11), as well as on relaxation times (12) of specific resonances of the ligand.It has been shown that in neutral aqueous solution the pyridine coenzymes, DPN+ and DPNH, exist in rapid equilibrium between open and folded conformations (13-15). Investigations regarding the actual conformations of these coenzymes in the active sites of dehydrogenases showed discrepancies depending on the methodology used. The fluorescence energy-transfer study of Velick indicates that the reduced coenzyme appears to exist in an open conformation upon binding to most dehydrogenases; 3-phosphoglyceraldehyde dehydrogenase being the only exception (16,17). From a 5A x-ray map, Adams et al. (18) 1593 the reduced coenzyme exists in an intramolecular folded conformation in the active site of this enzyme (19). The observation of the upfield shift of both H8 and H2 protons on the adenine moiety of DPNH in the presence of this enzyme led to the above conclusions.We have recently studied again the binding of pyridine coenzymes to various dehydrogenases under various experimental conditions. From the results of our detailed study, we have been able to ascertain the conformation of coenzymes when bound to dehydrogenases in solution. MATERIALS AND METHODSAlcohol dehydrogenase from yeast (YADH) was purchased from both Calbiochem, Calif. and Sigma Chemical Corp., Mo. Lactate dehydrogenase from chicken muscle (M4LDH) and chicken heart (H4LDH) were prepared in our laboratory by Mr. Francis Stolzenbach (20). Lactate dehydrogenase from lobster tail was isolated and purified as reported elsewhere (21,22). The specific activities of dehydrogenases used in this study were measured before each experiment.

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Effects of environment on the folding of nicotinamide-adenine dinucleotides in aqueous solutions

Cited by 44 publications

References 15 publications

NADH Oxidase Activity of Indoleamine 2,3-Dioxygenase

NADH Oxidase Activity of Indoleamine 2,3-Dioxygenase

Dynamic and static quenching of 1,N6-ethenoadenine fluorescence in nicotinamide 1,N6-ethenoadenine dinucleotide and in 1,N6-etheno-9-(3-(indol-3-yl) propyl) adenine.

Conformations of Diphosphopyridine Coenzymes upon Binding to Dehydrogenases

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