0000000000311402
AUTHOR
Carolin Dreher
SDS-facilitated in vitro formation of a transmembrane B-type cytochrome is mediated by changes in local pH.
Abstract The folding and stabilization of α-helical transmembrane proteins are still not well understood. Following cofactor binding to a membrane protein provides a convenient method to monitor the formation of appropriate native structures. We have analyzed the assembly and stability of the transmembrane cytochrome b 559 ′, which can be efficiently assembled in vitro from a heme-binding PsbF homo-dimer by combining free heme with the apo-cytochrome b 559 ′. Unfolding of the protein dissolved in the mild detergent dodecyl maltoside may be induced by addition of SDS, which at high concentrations leads to dimer dissociation. Surprisingly, absorption spectroscopy reveals that heme binding and…
Fe- but not Mg-protophorphyrin IX binds to a transmembrane b-type cytochrome.
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 Mg(2+) 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 …