Time-resolved infrared spectroscopy of intermediates and products from photolysis of 1-(2-nitrophenyl)ethyl phosphates: Reaction of the 2- nitrosoacetophenone byproduct with thiols

Article abstract of DOI:10.1021/ja964430u

Rapid scan Fourier transform infrared (FTIR) spectroscopy and time- resolved single wavelength infrared (IR) spectroscopy have been applied to study the mechanism of photochemical release of adenosine 5'-triphosphate (ATP) from its P³-[1-(2-nitrophenyl)ethyl] ester (caged ATP). Bands arising from phosphate and non-phosphate vibrations characteristic of the aci-nitro anion intermediate and from free ATP and hyproducts of the nitrophenylethyl group have been assigned using ¹³C, ¹⁵N, and ¹⁸O isotopomers of caged ATP. Monitoring several of these bands using time-resolved single frequency IR spectroscopy confirms that release of ATP occurs in a single exponential process synchronous with the decay of the aci-nitro anion intermediate. Spectral characteristics of the reaction products arising from the l-(2- nitrophenyl)ethyl group in the absence and presence of dithiothreitol (DTT) have been determined. The major final byproduct from photolysis conducted in the presence of DTT is 3-methylanthranil. The mechanism of formation of this compound from 2-nitrosoacetophenone has been investigated in detail by a combination of spectroscopic, kinetic, and chemical methods and reconciled with earlier data. The byproduct species likely to be present on the time scale of most biological experiments using caged compounds is 2- hydroxylaminoacetophenone as a mixture of ring-chain tautomers.