Sampling field heterogeneity at the heme of c-type cytochromes by spectral hole burning spectroscopy and electrostatic calculations

Monique Laberge, Martin Köhler, Jane M. Vanderkooi, Josef Friedrich

Research output: Contribution to journalArticlepeer-review

24 Scopus citations

Abstract

We report on a comparative investigation of the heme pocket fields of two Zn-substituted c-type cytochromes - namely yeast and horse heart cytochromes c - using a combination of hole burning Stark spectroscopy and electrostatic calculations. The spectral hole burning experiments are consistent with different pocket fields experienced at the hemes of the respective cytochromes. In the case of horse heart Zn-cytochrome c, two distinguishable electronic origins with different electrostatic properties are observed. The yeast species, on the other hand, displays a single electronic origin. Electrostatic calculations and graphics modeling using the linearized finite-difference Poisson-Boltzmann equation performed at selected time intervals on nanosecond-molecular dynamics trajectories show that the hemes of the respective cytochromes sample different potentials as they explore conformational space. The electrostatic potentials generated by the protein matrix at the heme show different patterns in both cytochromes, and we suggest that the cytochromes differ by the number of 'electrostatic substates' that they can sample, thus accounting for the different spectral populations observed in the two cytochromes.

Original languageEnglish
Pages (from-to)3293-3304
Number of pages12
JournalBiophysical Journal
Volume77
Issue number6
DOIs
StatePublished - Dec 1999

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