Nitrophenylethyl (NPE)- and hydroxyphenacyl (HPA)-caged nucleotides were employed in a time-resolved Fourier transform IR spectroscopy study on Ras-catalyzed guanosine triphosphate (GTP) hydrolysis. A fast kinetic component was observed following the photolysis of NPE-caged nucleotides in the NPE-GTP-Ras complex. However, this kinetic component was not observed in the HPA-GTP-Ras experiment. This fast kinetic component was likely due to a chemical reaction between Ras and the detached caging group, nitrosoacetophenone. This communication serves as a note of caution in interpreting spectral changes and kinetic behavior of the enzymatic systems employing NPE-caged compounds.
Copyright 2001 John Wiley & Sons, Inc. Biopolymers (Biospectroscopy) 62: 147-149, 2001