The Complete Amino Acid Sequence of Subunit d of Rat Liver Mitochondrial H+-ATP Synthase1

Abstract

Subunit d of H(+)-ATP synthase from rat liver mitochondria was isolated from the purified enzyme by reverse-phase high performance liquid chromatography. The partial amino acid sequence of the subunit was determined by automated Edman degradation of the peptide fragments. The nucleotide sequence of subunit d of rat liver H(+)-ATP synthase was determined from a recombinant cDNA clone isolated by screening a rat hepatoma cell line H4TG cDNA library with a probe DNA. The sequence was composed of 581 nucleotides including a coding region for the import precursor of subunit d and noncoding regions on the 5'- and 3'- sides. The possible precursor of subunit d and its mature polypeptide deduced from the open reading frame consisted of 161 and 160 amino acid residues with molecular weights of 18,763 and 18,631, respectively. Subunit d is a hydrophilic protein with an isoelectric point of 6.19. The sequence of the rat subunit d is highly homologous with that of subunit d of bovine heart and slightly similar to that of the subunit d of the yeast mitochondria. However, it had no homology with the sequence of any of the subunits of bacterial or chloroplast H(+)-ATP synthase.

Synchronized transcriptional gene expression of H+-ATP synthase subunits in different tissues of Fischer 344 rats of different ages

“…As described previously [20], the probes used were a XhoI± BglII fragment of the cDNA for subunit b [21], an AccI±AvaI fragment of cDNA for subunit d [22], a HindIII±HindIII fragment of cDNA for subunit e [23], a HindIII±ApaLI fragment of cDNA for OSCP [24], a HindIII±ApaLI fragment of cDNA for IF1 [24], a HindIII±HindIII fragment of cDNA for F6 [25], a…”

Gene Expression of Subunit c(P1), Subunit c(P2), and Oligomycin Sensitivity-conferring Protein May Play a Key Role in Biogenesis of H+-ATP Synthase in Various Rat Tissues

“…1, the probes used were a XhoI-BglII fragment of cDNA for subunit b (13), an AccI-AvaII fragment of cDNA for subunit d (17), a HindIII-HindIII fragment of cDNA for subunit e (15), a HindIII-ApaLI fragment of cDNA for OSCP (16), a HindIII-ApaLI fragment of cDNA for IF1 (16), a HindIII-HindIII fragment of cDNA for F 6 (14), a XhoI-EcoRI fragment of cDNA for subunit c(P1) (16), and an AccI-EcoO1091 fragment of cDNA for subunit c(P2) (16). The probe for ␤-subunit mRNA was a fragment of 634 -925 bp of ␤-subunit cDNA (21) that was synthesized by a nested reverse transcription-polymerase chain reaction method using four primers (outer primers: 5Ј-ATAAGGTTGTGGATCTGCTGGCC-3Ј and 5Ј-AACTCAGCAATAGCACGGGACAGCA-3Ј; inner primers: 5Ј-GGGATATCGCGTTGGTATATGGGCAGATGA-3Ј and 5Ј-GGGCGGC-CGCACATAGATAGCCTGCACTGAG-3Ј).…”

“…To clarify the coordinate transcriptional regulatory system(s) of the mammalian H ϩ -ATP synthase, we have developed a simple and rapid purification method of H ϩ -ATP synthase from rat (8,9) that can carry out the physiological experiments. We then determined the primary sequences of the synthase subunits by the protein sequence and cDNA cloning techniques (13)(14)(15)(16)(17).…”

“…We then determined the primary sequences of the synthase subunits by the protein sequence and cDNA cloning techniques (13)(14)(15)(16)(17).…”

Functional Disorders of the Oxidative Phosphorylation System in the Heart Mitochondria of Mice with Juvenile Visceral Steatosis.

“…The mammalian H ϩ -ATP synthase is a supramolecule composed of at least 14 subunits 21,[24][25][26][27] that have a constant stoichiometry. 28,29) Six of them construct the catalytic site of ATP synthesis called Fig.…”