Infrared Spectroscopy of Nicotinamide Adenine Dinucleotides in One and Two Dimensions

Infrared Spectroscopy of Nicotinamide Adenine

Dinucleotides in One and Two Dimensions

Niall Simpson,a Daniel J. Shaw,a, b Pim W.J.M. Frederix, a Audrey H. Gillies,a Katrin

Adamczyk, a Gregory M. Greetham, c Michael Towrie, c Anthony W. Parker, c Paul A.

Hoskisson, b and Neil T. Hunt a* a) Department of Physics, University of Strathclyde, SUPA, 107 Rottenrow East, Glasgow,

G4 0NL, UK b) Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, 161

Cathedral Street, Glasgow, G4 0RE, UK c) Central Laser Facility, Research Complex at Harwell, Rutherford Appleton Laboratory,

Didcot, Oxon, OX11 0QX, UK Supporting Information:

Effect of solvent composition on the FTIR spectrum of NAD:

Figure S1: FTIR spectra of (a) NAD+ and (b) NADH at neutral pD in water and in a water-methanol d-4 mixture of volume ratio 30:70 (water: methanol). NMR1-4 and UV-vis5,6 studies have demonstrated the effect of unfolding of NAD+ and NADH with decreasing solvent polarity. Significant changes in the proton shifts were reported from ~10% methanol inclusion, with the unfolding effect reported to be saturated at 70%. Changes to the IR spectra of the dinucleotides here however show only small frequency shifts (maximum ~5cm -1) that likely arise from the altered solvent environment, rather than an unfolding process. Acid-induced disruption of the nicotinamide ring of NADH:

Figure S2: NMR spectra of NADH at pD 2.5-7 near chemical shift values of (a) 7 ppm and (b) 1-3 ppm. Peaks are assigned according to the numbering scheme introduced in Fig1 (main text). Peaks identified with asterisks indicate new transitions attributable to tetrahydronicotinamide species formed under mildly acidic conditions

(pH <5). 7 These can clearly be seen to increase in magnitude as peaks due to NADH decrease. Vibrational Relaxation of NAD:

Figure S3: Ultrafast pump-probe spectra of (a) NAD+, pD2.5 and (b) NADH, pD7. Values shown correspond to the vibrational lifetime for each of the major modes obtained via fitting of peak amplitudes to a single exponential function. References:

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