Wechselwirkungen der nichtfunktionellen Coenzymbindungsstelle in Dehydrogenasen mit [Nicotinamid‐ribofuranosyl]‐[ω‐(adenin‐9‐yl)‐n‐alkyl]‐pyrophosphaten

Jeck, Reinhard; Wilhelm, G.

doi:10.1002/jlac.197319730317

Cited by 8 publications

(4 citation statements)

References 32 publications

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“…While the fluorescence lifetime of ENAD+ is 9.2% of the value for EAMP, the relative quantum yield is only 5% of that observed after snake venom treatment, taking into account a correction for hypochromism in the dinucleotide, an observation which is also consistent with a folded structure (22,(37)(38)(39)(40)(41)(42).…”

mentioning

confidence: 79%

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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confidence: 79%

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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“…The syntheses of the nicotinamide riboside w-(adenin-9yl)-n-alkyl diphosphate analogues were described previously (Jeck & Wilhelm, 1973), as were the syntheses of the nicotinamide riboside u>-(4-acetylanilino)-«-alkyl diphosphate analogues (Vutz et al, 1980). NMN was produced by hydrolyzing NAD"1" according to Jeck et al (1974).…”

Section: Methodsmentioning

confidence: 99%

“…NMN was produced by hydrolyzing NAD"1" according to Jeck et al (1974). These analogues and NMN were reduced to the dihydro forms by treatment with sodium dithionite at pH 8 (Jeck & Wilhelm, 1973) and purified by chromatography on a Sephadex G-10 column (2 X 200 cm) developed with sodium glycine buffer, pH 9.3, at 10 mL/h. The reduced coenzyme was completely separated from byproducts and, in the case of the adenine alkyl analogues, was identified by an absorption spectrum with maxima at 260 and 340 nm ( = 14000 and 6200 M™1 cm™1, respectively).…”

Section: Methodsmentioning

confidence: 99%

Kinetics and native and modified liver alcohol dehydrogenase with coenzyme analogs: isomerization of enzyme-nicotinamide adenine dinucleotide complex

Plapp¹,

Sogin²,

Dworschack³

et al. 1986

Biochemistry

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Coenzyme analogues with the adenosine ribose replaced with n-propyl, n-butyl, and n-pentyl groups; coenzyme analogues with the adenosine replaced with 3-(4-acetylanilino)propyl and 6-(4-acetylanilino)hexyl moieties; and nicotinamide mononucleotide, nicotinamide hypoxanthine dinucleotide, and 3-acetylpyridine adenine dinucleotide were used in steady-state kinetic studies with native and activated, amidinated enzymes. The Michaelis and inhibition constants increased up to 100-fold upon modification of coenzyme or enzyme. Turnover numbers with NAD+ and ethanol increased in some cases up to 10-fold due to increased rates of dissociation of enzyme-reduced coenzyme complexes. Rates of dissociation of oxidized coenzyme appeared to be mostly unaffected, but the values calculated (10-60 s-1) were significantly less than the turnover numbers with acetaldehyde and reduced coenzyme (20-900 s-1, at pH 8, 25 degrees C). Rates of association of coenzyme analogues also decreased up to 100-fold. When Lys-228 in the adenosine binding site was picolinimidylated, turnover numbers increased about 10-fold with NAD(H). Furthermore, the pH dependencies for association and dissociation of NAD+ and turnover number with NAD+ and ethanol showed the fastest rates above a pK value of 8.0. Turnover with NADH and acetaldehyde was fastest below a pK value of 8.1. These results can be explained by a mechanism in which isomerization of the enzyme-NAD+ complex (110 s-1) is partially rate limiting in turnover with NAD+ and ethanol (60 s-1) and is controlled by ionization of the hydrogen-bonded system that includes the water ligated to the catalytic zinc and the imidazole group of His-51.

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“…For example, nicotinamideribose-diphospho-5-pentyl-adenine, where the adenine ribose is replaced by a pentamethylene chain, is an active coenzyme analogue for horse liver alcohol dehydrogenase with a V larger than that observed with NAD' [23].…”

Section: Discussionmentioning

confidence: 99%

X-Ray Studies of the Binding of Cibacron Blue F3GA to Liver Alcohol Dehydrogenase

et al. 1979

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Wechselwirkungen der nichtfunktionellen Coenzymbindungsstelle in Dehydrogenasen mit [Nicotinamid‐ribofuranosyl]‐[ω‐(adenin‐9‐yl)‐n‐alkyl]‐pyrophosphaten

Cited by 8 publications

References 32 publications

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.

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.

Kinetics and native and modified liver alcohol dehydrogenase with coenzyme analogs: isomerization of enzyme-nicotinamide adenine dinucleotide complex

X-Ray Studies of the Binding of Cibacron Blue F3GA to Liver Alcohol Dehydrogenase

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