Primary structure, import, and assembly of the yeast homolog of succinate dehydrogenase flavoprotein

• Published in: Proceedings of the National Academy of Sciences - PNAS 1992-09, Vol.89 (17), p.8011-8015
• Main Authors: Schulke, N. (Howard Hughes Medical Institute, New York, NY), Blobel, G, Pain, D
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Amino acids; assembly; Base Sequence; Biological and medical sciences; Biological Transport; Carbonates; Cell Compartmentation; Ceres; CLONACION; CLONAGE; Cloning, Molecular; Enzymes; Fundamental and applied biological sciences. Psychology; Generally accepted auditing standards; Genes; Genes, Fungal; GENETICA MOLECULAR; GENETIQUE MOLECULAIRE; Growth, nutrition, metabolism, transports, enzymes. Molecular biology; Imports; LEVADURA; LEVURE; Membrane proteins; Microbiology; Mitochondria; Mitochondria - enzymology; MITOCHONDRIE; MITOCONDRIA; Molecular Sequence Data; Mycology; NUCLEOTIDE; nucleotide sequence; NUCLEOTIDOS; Oligodeoxyribonucleotides - chemistry; Polymerase Chain Reaction; predictions; Protein Precursors - metabolism; PROTEINAS; PROTEINE; Proteins; SACCHAROMYCES CEREVISIAE; Sequence Alignment; succinate dehydrogenase; Succinate Dehydrogenase - chemistry; Succinate Dehydrogenase - metabolism; SUCCINATE DESHYDROGENASE; SUCINATO DESHIDROGENASA; Yeasts
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.89.17.8011

We have isolated a homolog for the flavoprotein subunit of succinate dehydrogenase [succinate:(acceptor) oxidoreductase, EC 1.3.99.1] from Saccharomyces cerevisiae and used the obtained peptide sequences to clone and characterize the corresponding gene. It contained an open reading frame of 1923 base pairs and encoded a protein of 640 amino acids (Mr, 70,238) that showed approximately 49% and approximately 28% identity with the Escherichia coli and Bacillus subtilis enzymes, respectively. All features of the FAD cofactor binding site were completely conserved. Comparison of the deduced protein sequence with the N-terminal sequence determined from the isolated protein revealed an N-terminal extension of 28 amino acids that presumably represents a mitochondrial signal sequence. After in vitro transcription and translation, the preprotein was efficiently imported into isolated yeast mitochondria, cleaved to its mature form, and assembled into the membrane-bound succinate dehydrogenase complex