Transmembrane topology of the Rieske Fe/S protein of the cytochrome b6/f complex from spinach chloroplasts.
The topology of the Rieske protein of the cytochrome b6/f complex in thylakoids from spinach chloroplasts was examined by protease protection experiments as well as polypeptide extraction assays using solutions of chaotropic salts or alkaline pH. While neither thermolysin nor trypsin cleave any of the Rieske protein when added to the stromal side of the thylakoid membrane, proteinase K is capable of removing approximately four residues from its NH2-terminus. The protein is resistant to membrane extraction by 0.1 M Na2CO3 or 2 M NaBr but is quantitatively released by 0.1 M NaOH. Treatment of thylakoids with 2 M NaSCN leads to extraction of variable amounts of the protein, depending on the presence or absence of sucrose in the medium which apparently stabilizes the cytochrome complex. From these results we conclude that the Rieske protein is an integral component of the cytochrome complex which spans the thylakoid membrane with a single hydrophobic segment and is anchored predominantly by electrostatic interactions.
Karnauchov
I
I
Botanisches Institut der Ludwig-Maximilians-Universität, Munich, Germany.
Herrmann
R G
RG
Klösgen
R B
RB
D015153:Blotting, Western; D001965:Bromides / Q000494:pharmacology; D002254:Carbonates / Q000494:pharmacology; D002736:Chloroplasts / Q000737:chemistry*; D003573:Cytochrome b Group / Q000737:chemistry*; D045346:Cytochrome b6f Complex; D014450:Electron Transport Complex III; D019286:Endopeptidase K / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007506:Iron-Sulfur Proteins / Q000737:chemistry* / Q000378:metabolism; D008565:Membrane Proteins / Q000737:chemistry* / Q000378:metabolism; D010940:Plant Proteins / Q000737:chemistry / Q000378:metabolism; D011487:Protein Conformation; D017670:Sodium Compounds / Q000494:pharmacology; D012972:Sodium Hydroxide / Q000494:pharmacology; D018724:Spinacia oleracea / Q000737:chemistry; D013696:Temperature; D013820:Thermolysin / Q000378:metabolism; D013861:Thiocyanates / Q000494:pharmacology; D014357:Trypsin / Q000378:metabolism
D001965:Bromides; D002254:Carbonates; D003573:Cytochrome b Group; D007506:Iron-Sulfur Proteins; D008565:Membrane Proteins; D010940:Plant Proteins; C037989:Rieske iron-sulfur protein; D017670:Sodium Compounds; D013861:Thiocyanates; C005686:sodium carbonate; D012972:Sodium Hydroxide; C024553:sodium thiocyanate; D045346:Cytochrome b6f Complex; D014357:Trypsin; D019286:Endopeptidase K; D013820:Thermolysin; D014450:Electron Transport Complex III; C027938:sodium bromide