Complete amino acid sequence of subunit e of rat liver mitochondrial hydrogen ion-ATP synthase

Higuti, Tomihiko; Kuroiwa, Kayo; Kawamura, Yoshihiro; Yoshihara, Yutaka

doi:10.1021/bi00164a022

Cited by 25 publications

(14 citation statements)

References 35 publications

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“…The COOHterminal peptide of bovine e against which our anti-e antibodies were raised has the amino acid sequence ERELAEAQEDTILK. The corresponding segment of e from rat liver has the sequence ERELAEAEDVSIFK (24). As seen in Fig.…”

Section: Resultsmentioning

confidence: 99%

Membrane Topography and Near-neighbor Relationships of the Mitochondrial ATP Synthase Subunits e, f, and g

Belogrudov

Tomich

Hatefi

1996

Journal of Biological Chemistry

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The well characterized subunits of the bovine ATP synthase complex are the ␣, ␤, ␥, ␦, and ⑀ subunits of the catalytic sector, F 1 ; the ATPase inhibitor protein; and subunits a, b, c, and d, OSCP (oligomycin sensitivityconferring protein), F 6 , and A6L, which are present in the membrane sector, F 0 , and the 45-Å-long stalk that connects F 1 to F 0 . It has been shown recently that bovine ATP synthase preparations also contain three small polypeptides, designated e, f, and g, with respective molecular masses of 8.2, 10.2, and 11.3 kDa. To ascertain their involvement as bona fide subunits of the ATP synthase and to investigate their membrane topography and proximity to the above ATP synthase subunits, polyclonal antipeptide antibodies were raised in the rabbit to the COOH-terminal amino acid residues 57-70 of e, 75-86 of f, and 91-102 of g. It was shown that (i) e, f, and g could be immunoprecipitated with anti-OSCP IgG from a fraction of bovine submitochondrial particles enriched in oligomycin-sensitive ATPase; (ii) the NH 2 termini of f and g are exposed on the matrix side of the mitochondrial inner membrane and can be curtailed by proteolysis; (iii) the COOH termini of all three polypeptides are exposed on the cytosolic side of the inner membrane; and (iv) f cross-links to A6L and to g, and e crosslinks to g and appears to form an e-e dimer. Thus, the bovine ATP synthase complex appears to have 16 unlike subunits, twice as many as its counterpart in Escherichia coli.

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Section: Resultsmentioning

confidence: 99%

Membrane Topography and Near-neighbor Relationships of the Mitochondrial ATP Synthase Subunits e, f, and g

Belogrudov

Tomich

Hatefi

1996

Journal of Biological Chemistry

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“…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…”

Section: P-labeled Probe Dnasmentioning

confidence: 99%

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

Himeda¹,

Morokami²,

Arakaki³

et al. 2000

Eur J Biochem

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Little is known about the relationship between the stoichiometry of polypeptides of multisubunit enzyme complexes and the absolute amount of each transcript of the complexes in mammalian tissues. Here we showed that the absolute amounts of the transcripts of most subunits of rat H 1 -ATP synthase examined greatly differed in the different tissues, showing the following hierarchy of tissue-specificity: heart . kidney . brain < liver. However, surprisingly, there was no difference in the expression pattern of these in terms of the molar ratio of each transcript, indicating a nearly similar stoichiometric expression pattern irrespective of tissue or age of the rat. Therefore, the present finding clearly indicates that most of the transcripts of the 16 subunits of rat H 1 -ATP synthase were concertedly and synchronously expressed, having a constant expression pattern of the transcripts, irrespective of tissue or age of the rats. This is the first report of the absolute amounts of the transcripts of this multisubunit enzyme.Keywords: H 1 -ATP synthase; expression pattern; transcript; stoichiometry; synchronization.The mechanism of the transcriptional regulation of the genes involved in the biosynthesis and assembly of multisubunit mitochondrial enzyme complexes, e.g. [14,17±18a]. Subunit a and A6L are encoded by the mitochondrial genome, and all of the other subunits are separately encoded by nuclear chromosomes; however, the relationship between the stoichiometry of polypeptides of the complexes and the absolute amount of each transcript of this multisubunit enzyme complex in mammalian tissues is still entirely unknown.In the present work, we determined the absolute amount of nine species of mRNAs for eight nucleus-encoded subunits of H 1 -ATP synthase in the brain, liver, heart, and kidney tissues of 2-week-old to 90-week-old male Fischer 344 rats by using highly purified poly(A) 1 RNA and the mRNAs synthesized by an in vitro transcriptional system. M A T E R I A L S A N D M E T H O D S AnimalsMale Fischer 344 rats were obtained from Charles River Japan Inc (Atsugi, Japan), and were genetically identical because of approximately 300 matings between sister and brother. Purification of Poly(A)1 RNAThe rats were decapitated, and tissues of brain, liver, heart, and kidney were rapidly removed. Total RNA from these tissues was isolated by the guanidinium thiocyanate method [19], followed by centrifugation at 120 000 g for 24 h in 5.7 m CsCl solution and then extraction with acid phenol. The poly(A) 1 RNA was purified with Oligotex-dT30 (Takara) according to the manufacturer's instructions, and the purification steps with Oligotex-dT30 were repeated twice. The poly(A) 1 RNA eluted from the 2nd Oligotex-dT30 was extracted with acid phenol, precipitated with 70% ethanol, dried, and dissolved in diethyl pyrocarbonate-treated water. The amount of poly(A) 1 RNA was determined from the absorbance at 260 nm (one unit of absorbance at 260 nm 40 mg´mL 21). The A260/A280 ratios were in the range 1.8±2.0. Contamination by ribosoma...

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“…In addition, other agents may also modify its activity. Higuti et al (7) have proposed that the e subunit contains a putative Ca 2ϩ binding site, suggesting that a Ca 2ϩ -e subunit complex participates in regulating e subunit binding or activity (7). Regulating F 1 F 0 -ATP synthase activity by altering transcriptional responses may seem less probable because the required enzyme machinery must be constantly responsive to changes in energy balance and substrate concentration.…”

Section: Figmentioning

confidence: 99%

“…Its activity is partly regulated by the availability of substrates and the proton flux through the membrane (1,2). The synthase complex consists of a soluble F 1 catalytic unit with the F 0 portion composed of a joining stalk and inner membrane proteins (1)(2)(3)(4)(5)(6)(7). Proton flux through F 0 membrane proteins has been postulated to cause conformational changes, which may pass to the catalytic F 1 subcomplex through the stalk (2).…”

mentioning

confidence: 99%

“…Their functions are unknown. In a screen for genes regulated by the level of dietary lipids and carbohydrates (8), we isolated and characterized low fat mammary 1 (Lfm1), 1 whose predicted protein sequence is 96% and 98% similar to e subunit sequences in bovine heart (4) and rat liver (7). Since the latter two proteins are 94% similar, we suggested that Lfm1 is the murine F 1 F 0 -ATP synthase e subunit gene (8).…”

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confidence: 99%

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The e Subunit Gene of Murine F1F0-ATP Synthase

Swartz

Park

Visek

et al. 1996

Journal of Biological Chemistry

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Genomic sequences encoding murine Lfm1, whose predicted protein sequence is 96% and 98% similar to bovine and rat F 1 F 0 -ATP synthase e subunits (respectively), have been amplified from BALB/cByJ DNA, cloned, and sequenced. The 1.1-kilobase gene has 3 introns and 4 exons, and its coding sequence differs by two nucleotides compared to the previously published BALB/cHnn Lfm1 cDNA sequence. A PstI restriction site polymorphism in intron 2 between C57BL/6J and Mus spretus was used to map this gene to Chromosome 5 near D5Mit9. Related sequences were mapped on Chromosomes 8, 11, and 2 unlinked loci on Chromosome 2 using Southern blot analyses with the 1.1-kilobase gene as probe. Previous studies from this laboratory indicated that the Lfm1/e subunit was regulated by the level of dietary fat and carbohydrate. Northern hybridization analyses demonstrated that e subunit mRNA abundance showed statistically significant differences (p < 0.025) between hearts of BALB/c mice fed 3% and those fed 20% corn oil for 2 weeks and in liver (p < 0.05) from the same animals. Significant differences were also observed in hepatic and heart mRNA expression at different times after eating in animals subjected to a fast/refeed regimen. The implications of the high degree of sequence similarity to the e subunit for rat and bovine F 1 F 0 -ATP synthase and its regulation by diet are discussed.

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Complete amino acid sequence of subunit e of rat liver mitochondrial hydrogen ion-ATP synthase

Cited by 25 publications

References 35 publications

Membrane Topography and Near-neighbor Relationships of the Mitochondrial ATP Synthase Subunits e, f, and g

Membrane Topography and Near-neighbor Relationships of the Mitochondrial ATP Synthase Subunits e, f, and g

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

The e Subunit Gene of Murine F1F0-ATP Synthase

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