Time-resolved FTIR study of CO recombination with horseradish peroxidase.

摘要

Vibrational changes associated with CO recombination to ferrous horseradish peroxidase were investigated by rapid-scan FTIR (Fourier-transform IR) spectroscopy in the 1200-2200 cm(-1) range. At pH 6.0, two conformers of bound CO are present that appear as negative bands at 1905 and 1934 cm(-1) in photolysis spectra. Their recombination rate constants are identical, confirming that they arise from two substates of bound CO that are in rapid thermal equilibrium, rather than from heterogeneous protein sites. A smaller positive band at 2134 cm(-1) also appears on photolysis and decays with the same rate constant, indicative of an intraprotein geminate site involved in recombination or, possibly, a weak-affinity surface CO-binding site. Other signals arising from protein and haem in the 1700-1200 cm(-1) range can also be time-resolved with similar kinetics.