Rapid events in protein folding studied by laser-induced pH and temperature jumps

Abstract

Traditional methods, such as stopped flow, used to study early events in protein folding are limited, by instrument dead times, to investigating events which occur milliseconds after the initiation of folding. We have developed a laser-based temperature jump apparatus capable of measuring changes in the fluorescence of proteins undergoing folding or unfolding on the microsecond timescale following a laser-induced pH, or temperature, jump. The development of the system is discussed and the results for experiments with RNase T1 and Apomyoglobin are summarized.