Heme binding properties of heterologously expressed spinach cytochrome b(6): implications for transmembrane b-type cytochrome formation.
In vivo and in vitro requirements for the formation of cytochrome b(6) were examined to analyze the mechanisms of transmembrane b-type cytochrome formation. After heterologous expression of spinach cytochrome b(6), formation of the holo-cytochrome was observed within the E. coli inner membrane. The transmembrane orientation of cytochrome b(6) appeared not to be critical for heme binding and holo-cytochrome formation. Furthermore, in vitro reconstitution of cytochrome b(6) was possible under oxidizing as well as under reducing conditions. Taken together these observations strongly indicate that transmembrane b-type cytochromes can spontaneously assemble in vitro as well as in a membrane.
Dreher
Carolin
C
Institut für Biochemie und Molekularbiologie, Zentrum für Biochemie und Molekulare Zellforschung, Albert-Ludwigs-Universität Freiburg, Stefan-Meier-Strasse 19, Freiburg, Germany.
Prodöhl
Alexander
A
Weber
Mathias
M
Schneider
Dirk
D
D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D045347:Cytochromes b6 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D005816:Genetic Complementation Test; D006418:Heme / Q000378:metabolism*; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D010084:Oxidation-Reduction; D033902:Periplasmic Binding Proteins / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013056:Spectrophotometry, Ultraviolet; D018724:Spinacia oleracea / Q000378:metabolism*
D045347:Cytochromes b6; D029968:Escherichia coli Proteins; C461619:MalE protein, E coli; D008565:Membrane Proteins; D033902:Periplasmic Binding Proteins; D011993:Recombinant Fusion Proteins; D006418:Heme