Conformational Dynamics of Cytochrome P-450cam As Monitored by Photoacoustic Calorimetry

Carmelo Di Primo, Gaston Hui Bon Hoa, Eric Deprez, Pierre Douzou, Stephen G. Sligar

Research output: Contribution to journalArticlepeer-review

Abstract

Conformational transitions of cytochrome P-450cam following the dissociation of CO from the ferrous heme were investigated by using photoacoustic calorimetry. The effect of substrate association on the acoustic signal was also examined. Results show that the conformational dynamics of cytochrome P-450cam substrate-free protein occur faster than 10 ns, which is the time scale of the instrument response. The enthalpy and volume change for the dissociation reaction are 2.2 kcal mol-1 and 1.8 mL mol-1, respectively. Upon addition of camphor, the reaction is markedly slowed. An intermediate is formed whose lifetime is 130 ns at 17 °C. The overall enthalpy and volume changes are −15.9 kcal mol-1 and 10.3 mL mol-1, respectively. These results, together with published transient Raman spectra [Wells, A. V., Pusheng, L., Champion, P. M., Martinis, S. A., & Sligar, S. G. (1992) Biochemistry 31, 4384-4393] suggest that camphor leaves the heme pocket concomitant with the photoinduced expulsion of CO into the solvent and induces a considerable conformational change in the protein.

Original languageEnglish (US)
Pages (from-to)3671-3676
Number of pages6
JournalBiochemistry
Volume32
Issue number14
DOIs
StatePublished - Apr 1 1993

ASJC Scopus subject areas

  • Biochemistry

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