The primary structure of the iron-sulfur subunit of ubiquinol-cytochrome c reductase from Neurospora, determined by cDNA and gene sequencing.
The primary structure of the iron-sulfur subunit of ubiquinol-cytochrome c reductase from Neurospora mitochondria was determined by cDNA and genomic DNA sequencing. A first cDNA was identified from a cDNA bank cloned in Escherichia coli by hybridization selection of mRNA, cell-free protein synthesis and immunoadsorption. Further cDNA and geonomic DNA were identified by colony filter hybridization. The N-terminal sequence of the mature protein was determined by automated Edman degradation. From the sequence a molecular mass of 24749 Da results for the precursor protein and of 21556 Da for the mature protein. The presequence consists of 32 amino acids with four arginines as the only charged residues. The mature protein consists of 199 amino acids. It is characterized by a small N-terminal hydrophilic part of 29 residues, a hydrophobic stretch of 25 residues and a large C-terminal hydrophilic domain of 145 residues. The only four cysteines of the protein, which are assumed to bind the 2 Fe-2S cluster, are located in a moderate hydrophobic region of this large domain. Cysteines 3 and 4 are unusually arranged in that they are separated by only one proline. From sequence data the arrangement of the subunit in the membrane is deduced.
Harnisch
U
U
Weiss
H
H
Sebald
W
W
D000595:Amino Acid Sequence; D001483:Base Sequence; D055598:Chemical Phenomena; D002621:Chemistry; D003001:Cloning, Molecular; D004247:DNA; D014450:Electron Transport Complex III; D005796:Genes; D007506:Iron-Sulfur Proteins / Q000235:genetics / Q000302:isolation & purification*; D008667:Metalloproteins / Q000302:isolation & purification*; D009097:Multienzyme Complexes / Q000235:genetics / Q000302:isolation & purification*; D009247:NADH, NADPH Oxidoreductases / Q000302:isolation & purification*; D009491:Neurospora / Q000201:enzymology*; D009693:Nucleic Acid Hybridization; D011808:Quinone Reductases / Q000235:genetics / Q000302:isolation & purification*
D007506:Iron-Sulfur Proteins; D008667:Metalloproteins; D009097:Multienzyme Complexes; D004247:DNA; D009247:NADH, NADPH Oxidoreductases; D011808:Quinone Reductases; D014450:Electron Transport Complex III