Fe- but not Mg-protophorphyrin IX binds to a transmembrane b-type cytochrome

Abstract

Transmembrane b-type cytochromes, which are crucially involved in electron transfer chains, bind one or more heme (Fe-protoporphyrin IX) molecules non-covalently. Similarly, chlorophylls are typically also non-covalently bound by several membrane integral polypeptides involved in photosynthesis. While both, chlorophyll and heme, are tetrapyrrole macrocycles, they have different substituents at the tetrapyrrole ring moiety. Furthermore, the central metal ion is Mg²⁺ in chlorophyll and Fe^2+/3+ in heme. As heme and chlorophyll a have similar structures and might both be ligated by two histidine residues of a polypeptide chain, and as the local concentration of chlorophyll a might be up to 100-times higher than the concentration of heme, the question arises, as to how an organism ensures specific binding of heme, but not of chlorophyll, to transmembrane apo-cytochromes involved in photosynthetic electron transfer reactions. As shown here, Fe-protoporphyrin IX derivatives with modified substituents at the tetrapyrrole ring moiety still bind to an apo-cytochrome; however, association appears to be reduced. This indicates that hydrophobic and polar interactions of the ring substituents with the protein moiety stabilize the protein/heme-complex but are not essential per se. However, removal or replacement of the central Fe-ion completely abolishes formation of a holo-protein complex, and thus the central iron ion appears to determine heme binding to apo-cytochrome b₆.

• Assembly
• heme
• membrane protein

Acknowledgements

This work was supported by grants from the Ministry of Science, Research and Arts of Baden-Württem-berg, from the Centre of Complex Matter and from the University of Mainz. We thank Daniel Otzen for helpful discussions and Hildegard Pearson for critically reading the manuscript.