Cytochromes c' in their reaction with ethyl isocyanide.
The binding of ethyl isocyanide (EIC) to a representative number of cytochromes c' is demonstrated. Spectroscopic and equilibrium constants have been measured and compared for the binding of EIC to cytochromes c' from the photosynthetic bacteria Chromatium vinosum, Rhodopseudomonas palustris, Rhodospirillum rubrum, and Rhodopseudomonas sphaeroides. While the absorption spectra of the EIC complexes resemble those of EIC complexes of other high-spin hemoproteins, the Soret half band widths and extinction coefficients per heme exhibit more than a 2-fold difference with the values of C. vinosum being most similar to those of Rh. sphaeroides and of Rh. palustris similar to those of Rs. rubrum. The cytochromes exhibit binding equilibria consistent with the ligation of one molecule of EIC per heme in contrast to the reported binding of more than one molecule of CO per heme. The binding constants exhibit more than a 1000-fold difference with the values of C. vinosum being closely similar to those of Rh. sphaeroides and of Rh. palustris similar to those of Rs. rubrum. The lack of correlation between EIC and CO binding properties indicates that electronic factors do not determine the difference in EIC binding properties. The observed correlation between the extinction coefficients, half band widths, and equilibrium constants for EIC complex formation provides the first spectroscopic evidence that the differences in binding properties are associated with sterically hindered ligation to the heme. Although the differences in binding properties provide evidence of steric hindrance, the EIC binding constants for particular cytochromes c' indicate that the distal heme binding site is more accessible than previously indicated.(ABSTRACT TRUNCATED AT 250 WORDS)
D000595:Amino Acid Sequence; D002844:Chromatium / Q000378:metabolism*; D003574:Cytochrome c Group / Q000378:metabolism*; D007700:Kinetics; D009570:Nitriles / Q000378:metabolism*; D011485:Protein Binding; D012242:Rhodobacter sphaeroides / Q000378:metabolism*; D012241:Rhodopseudomonas / Q000378:metabolism*; D012247:Rhodospirillum rubrum / Q000378:metabolism*; D013045:Species Specificity
D003574:Cytochrome c Group; D009570:Nitriles; C001187:ethyl isocyanide