Raman spectroscopy of oxidized and reduced nicotinamide adenine dinucleotides

Abstract

We have measured the Raman spectra of oxidized nicotinamide adenine dinucleotide, NAD+, and its reduced form, NADH, as well as a series of fragments and analogues of NAD+ and NADH. In addition, we have studied the effects of pH as well as deuteration of the exchangeable protons on the Raman spectra of these molecules. In comparing the positions and intensities of the peaks in the fragment and analogue spectra with those of NADH and NAD+, we find that it is useful to consider these large molecules as consisting of component parts, namely, adenine, two ribose groups, two phosphate groups, and nicotinamide, for the purpose of assigning their spectral features. The Raman bands of NADH and NAD+ are found generally to arise from molecular motions in one or another of these molecular moieties, although some peaks are not quite so easily identified in this way. This type of assignment is the first step in a detailed understanding of the Raman spectra of NAD+ and NADH. This is needed to understand the binding properties of NADH and NAD+ acting as coenzymes with the NAD-linked dehydrogenases as deduced recently by using Raman spectroscopy.

A study of the binding of NADP coenzymes to dihydrofolate reductase by raman difference spectroscopy

“…This band is not present in the spectra of NAD cofactors and derivatives, like ADP (Yue et al, 1986) or in the spectrum of NADH bound to DHFR (in ternary complex with trimethoprim; data not shown). Moreover, model compound studies on glucose 6-phosphate show a band at this position (977 cm-') which shifts down 42 cm-' upon '80-labelling of the phosphate group (data not shown) ; this band is assigned to the P = 0 stretch of the phosphate ester.…”

“…We have labelled peaks as either adenine (A), nicotinamide (N), ribose (R), phosphate (P), or diphosphate (PP) in the data of Fig. 1 for those peaks that are localized on these groups (Yue et al, 1986). We are able to make a somewhat more precise identification of solution bands (see Yue et al, 1986).…”

NMR Spectra and Potentiometry Studies of Aluminum(III) Binding with Coenzyme NAD+ in Acidic Aqueous Solutions

“…7 (2) Two sharp peaks at -14.9 ppm and -15.1 ppm can probably be attributed to the monodentate binding Al-NAD + complex species. The difference between the chemical shifts of these species indicates that NAD + contain charged nitrogen in the pyridine ring, and that there is an electrostatic interaction between this nitrogen 17 The X-ray crystal structures of metal ion complexes with adenine phosphate have shown three kinds of binding sites for metal coordination: 6 (1) phosphate oxygen; (2) ribose sugar alcohol oxygen; and (3) adenine ring nitrogen. With the coordination abilities of NAD + to metal ions, all of the exposed nitrogen atoms of the adenine ring, except for the amino group, have been shown to be capable of σ-donor binding to metal ions, e.g.…”

“…They are composed of two 5′-nucleotide bases, AMP and nicotinamide ribose 5′-phosphate, which are linked together by a pyrophosphate bridge. 6 The structure of NAD + with its atom number scheme is shown in Fig. 1.…”

“…13,14 Also, for purine nucleotides, in general, metal-heterocyclic nitrogen binding at N7′ is preferred over N1 or N3 with metal ions such as Ca 2+ , Co 2+ , Ni 2+ , Pt 2+ , Cu 2+ , Zn 2+ , La 3+ and Eu 3+ . 6,[12][13][14]20 However, no study concerning the interaction between Al 3+ and NAD + has been found in the available literature. In the present work, we utilized potentiometry, multinuclear ( 1 H, 13 C, 27 Al, 31 P) and two-dimensional ( ) and 2:1 (AlL2 -) species, and dinuclear 2:2 (Al2L2 2+ ) species.…”