The chloroplastic protein import machinery contains a Rieske-type iron-sulfur cluster and a mononuclear iron-binding protein.
Transport of precursor proteins across the chloroplastic envelope membranes requires the interaction of protein translocons localized in both the outer and inner envelope membranes. Analysis by blue native gel electrophoresis revealed that the translocon of the inner envelope membranes consisted of at least six proteins with molecular weights of 36, 45, 52, 60, 100 and 110 kDa, respectively. Tic110 and ClpC, identified as components of the protein import apparatus of the inner envelope membrane, were prominent constituents of this complex. The amino acid sequence of the 52 kDa protein, deduced from the cDNA, contains a predicted Rieske-type iron-sulfur cluster and a mononuclear iron-binding site. Diethylpyrocarbonate, a Rieske-type protein-modifying reagent, inhibits the translocation of precursor protein across the inner envelope membrane, whereas binding of the precursor to the outer envelope membrane is still possible. In another independent experimental approach, the 52 kDa protein could be co-purified with a trapped precursor protein in association with the chloroplast protein translocon subunits Toc86, Toc75, Toc34 and Tic110. Together, these results strongly suggest that the 52 kDa protein, named Tic55 due to its calculated molecular weight, is a member of the chloroplastic inner envelope protein translocon.
Caliebe
A
A
Botanisches Institut, Christian-Albrechts-Universität, Am Botanischen Garten 1-9, D-24118 Kiel, Germany.
Grimm
R
R
Kaiser
G
G
Lübeck
J
J
Soll
J
J
Heins
L
L
D000595:Amino Acid Sequence; D001665:Binding Sites; D002352:Carrier Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification*; D002736:Chloroplasts / Q000378:metabolism*; D003001:Cloning, Molecular; D004047:Diethyl Pyrocarbonate / Q000494:pharmacology; D014450:Electron Transport Complex III; D007425:Intracellular Membranes / Q000187:drug effects / Q000378:metabolism; D033862:Iron-Binding Proteins; D007506:Iron-Sulfur Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification*; D008565:Membrane Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D018532:Pisum sativum; D011498:Protein Precursors / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D033863:Transferrin-Binding Proteins
D002352:Carrier Proteins; D033862:Iron-Binding Proteins; D007506:Iron-Sulfur Proteins; D008565:Membrane Proteins; D011498:Protein Precursors; D011994:Recombinant Proteins; C037989:Rieske iron-sulfur protein; D033863:Transferrin-Binding Proteins; D014450:Electron Transport Complex III; D004047:Diethyl Pyrocarbonate