.beta. Subunit of rat liver mitochondrial ATP synthase: cDNA cloning, amino acid sequence, expression in Escherichia coli and structural relationship to adenylate kinase

Garboczi, David N.; Fox, Arthur H.; Gerring, S L; Pedersen, Peter L.

doi:10.1021/bi00402a008

Cited by 77 publications

(37 citation statements)

References 47 publications

(49 reference statements)

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“…fragment of the rat ATPase /~-subunR eDNA in pUCI9 [10] C~OX activity was measured spectrophotomgtrically, the content of COX IV and of ATPase fl-subunits w.s determined by Western blot analysis using tissue homogenates.…”

Section: Methodsmentioning

confidence: 99%

Low content of mitochondrial ATPase in brown adipose tissue is the result of post‐transcriptional regulation

et al. 1991

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The mI~NA levels of ATPase/5, ATPase 6, cytochrome oxidase (COX) Vlb and COX I subunit~ were found to be 2.4~13.8-fold higher in brown adipose tissue (BAT) than in heart, skeletal muscle, brain and liver of mice. The comparison with tissue contents of ATPase and COX revealed that the sele~=tive, 5-1 l-fold reduction of ATPase in BAT is not caused by decreased transcription of ATPase genes. Likewise, the ATPase ]~ and COX Vlb mRNA levels in cultured brown adipocytes were also not influenced by norepinephrine, which activated the expression of the UCP gene by two orders of magnitude. The results indicate that the biosynthesis of mitochondrial ATPase in BAT is post-transcriptionally regulatm~l. lional events are responsible for the selective reduction of the ATPase content in BAT. MATERIALS AND METHODSFour-week-old mice of Balb/c outbred strain were maintained at room temperature, on a 12 h light/12 h dark cycle. When indicated, animals were exposed to 4°C prior to use. The animals were killed by decapitation and ti.ssue samples for RNA isolation were frozen in liquid nitrogen. For other assays, 5~ homog~.:ates [w/v) were prepared in 0.25 M sucrose, 10 mM Tris/HCI, I mM EDTA, pH 7.4, and filtered through a 250/Jm nylon screen.The stromal-vascular ceils from BAT were isolated and cultivated as previously [71. Experiments were performed with cells cultured for 7 days.Total RNA was isolated as described in [8] and checked for purity and integrity. For Northern blot analysis, 20Mg RNA aliqnots were denatured in the presence of glyoxaL separated by eleetrophoresis in 1.2% agaros¢ gel and transferred to a nylon membrane (Hybond-N, Amersham) according to standard procedures [9]. Hybridization was carried out overnight at 68°C in 5x Dcnhardt's solution, 5x SSC, 50 mM sodium phosphate, pH 6.5. 0. !% SDS and 250 pg/ml of herring sperm DNA. Blots were washed in 2× SSC, 0.1 ~b SDS (2x |5 min, room temp.) and 0.Ix SSC, 0.1% SDS (2× 15 rain, 55°C). Following autoradiography the blots were stripped for 60 rain at 95~C in 0. I SDS and sequentially re-hybridized with several DNA pr3bes.The eDNA probes were generously provided by the following investigators: 1.26 kb E.coR! fragment of the rat ATPase /~-subunR eDNA in pUCI9

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Section: Methodsmentioning

confidence: 99%

Low content of mitochondrial ATPase in brown adipose tissue is the result of post‐transcriptional regulation

et al. 1991

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“…Adenine nucleotide translocase isoform 1 (ANT 1 ) mRNA levels were detected using a 1.4-kb ANT 1 cDNA fragment cloned from a human skeletal muscle library (ATCC, Rockville, Maryland) (23,35). ␤F 1 -ATPase mRNA levels were detected using a 1.8-kb ␤F 1 -ATPase cDNA fragment cloned from a human HeLa cell line library (ATCC) (4,37). An inducible heat shock protein (HSP70-1) mRNA level was detected by a 1.7-kb cDNA fragment cloned from human hippocampus (ATCC) (17,21).…”

Section: Preparation Of Isolated Heartmentioning

confidence: 99%

Temperature Threshold and Preservation of Signaling for Mitochondrial Membrane Proteins during Ischemia in Rabbit Heart

Xue

Song

et al. 1998

Cryobiology

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Temperature modulates both myocardial energy requirements and production. We have previously demonstrated that myocardial protection induced by hypothermic adaptation preserves expression of genes regulating heat shock protein and the nuclear-encoded mitochondrial proteins, the adenine nucleotide translocator isoform 1 (ANT 1 ), and the ␤ subunit of F 1 -ATPase (␤F 1 -ATPase). This preservation is associated with a reduction in ATP depletion similar to that noted in cardioplegic arrested hearts preserved at a critical temperature (30°C) or below. We tested the hypothesis that expression of these genes may also be subject to this temperature threshold phenomenon. Isolated perfused rabbit hearts were subjected to ischemic cardioplegic arrest at 4, 30, or 34°C for 120 min. Cardiac function indices and steady-state mRNA levels for ANT 1 , ␤F 1 -ATPase, and HSP70-1 were measured prior to ischemia (B) and after 45 min of reperfusion. Cardiac function was significantly depressed in the 34°C group. Ischemia at 34°C reduced steady-state mRNA levels for ANT 1 and ␤F 1 -ATPase from B, but these levels were similarly preserved at 4 and 30°C. HSP70-1 levels were mildly elevated (fourfold) above B to similar levels at all three temperatures. These results indicate that mRNA expression for ANT 1 and ␤F 1 -ATPase is specifically preserved in a pattern consistent with the temperature threshold phenomenon. HSP70-1 expression is not influenced by ischemic temperature. Preservation of gene expression for these mitochondrial proteins implies that signaling for mitochondrial biogenesis or resynthesis is maintained after ischemic insult. © 1998 Academic PressKey Words: adaptation; ANT 1 ; ␤F 1 -ATPase; cardioplegia; HSP70-1; hypothermia; myocardial ischemia; myocardial reperfusion; threshold.Temperature protects ischemic myocardium (2,3,6,7,15,34,36,38) and modulates both myocardial energy requirements and production (27). The relationship between energy supply and demand in the cardioplegic arrested heart remains fairly constant in the temperature range between 4 and 30°C (27), which is frequently used during cardiac surgical procedures. The myocardial supply/demand ratio demonstrates a threshold phenomenon by deteriorating rapidly at temperatures above 30°C (27). Accelerated ATP depletion and inability to reestablish highenergy phosphate stores during reperfusion accompany this deterioration. Thus, 30°C represents a critical temperature point (39), which triggers acceleration of metabolic energy utilization rate above the energy production rate. Provision of glucose as substrate in the cardioplegia solution can partially ameliorate this metabolic imbalance by promoting anaerobic ATP production (27).Recently, we have demonstrated that hypothermic exposure at 30°C prior to warm ischemia preserves myocardial function and ATP stores after reperfusion (28). This response represents an adaptation, whereby the heart becomes tolerant to a second exposure of the same or alternative type of stress. Hypothermic exposure induces a metabolic dow...

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“…2B) as follows. Most mass peaks corresponded closely (<0.1% mass difference) to the masses of tryptic peptides (17 limit peptides, 4 partials) predicted by direct translation of the Fl-13-subunit cDNA sequence [20,21]. The remaining 10 mass peaks differed from predicted Fl-13-subunit tryptic peptides by 16 Da or 32 Da, suggesting the possibility of methionine oxidation [22].…”

Section: Identification Of An Enamel Cell 52 Kda Calcium-bindingmentioning

confidence: 99%

“…Together, these data characterized the Fl-~-subunit as a (macroscopically) low affinity and moderate capacity calcium-binding protein. [20,21,23]) is represented by the identity ratio, with an ambiguous call (in parentheses; the expected residue is given) excluded. Initial sequencing yields were 5-12 pmol.…”

Section: Calcium-binding Properties Of the Fl-~-subunitmentioning

confidence: 99%

Mitochondrial ATP synthase F₁‐β‐subunit is a calcium‐binding protein

1996

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Mitochondrial ATP synthase is responsive to changes in cytosolic calcium concentration, but the regulatory mechanisms are unclear. Here we identified a major 52 kDa calciumbinding protein in rat enamel cells as the mitochondrial ATP synthase Fl-~subunit. The Fl-~-subunit behaved as a low affinity and moderate capacity calcium-binding protein during comparative 45Ca overlay analyses. Equivalent behavior was shown by the Fl-[I-subunit from rat liver mitochondria, but not by the homologous Fl-a-subunit, supporting the specificity of calcium binding. Evidence that the catalytic Fr~-subuuit binds calcium specifically introduces new mechanistic possibilities for regulating ATP synthase, and thereby coordinating ATP production with demand for ATP-fuelled calcium pump activity.

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.beta. Subunit of rat liver mitochondrial ATP synthase: cDNA cloning, amino acid sequence, expression in Escherichia coli and structural relationship to adenylate kinase

Cited by 77 publications

References 47 publications

Low content of mitochondrial ATPase in brown adipose tissue is the result of post‐transcriptional regulation

Low content of mitochondrial ATPase in brown adipose tissue is the result of post‐transcriptional regulation

Temperature Threshold and Preservation of Signaling for Mitochondrial Membrane Proteins during Ischemia in Rabbit Heart

Mitochondrial ATP synthase F₁‐β‐subunit is a calcium‐binding protein

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.beta. Subunit of rat liver mitochondrial ATP synthase: cDNA cloning, amino acid sequence, expression in Escherichia coli and structural relationship to adenylate kinase

Cited by 77 publications

References 47 publications

Low content of mitochondrial ATPase in brown adipose tissue is the result of post‐transcriptional regulation

Low content of mitochondrial ATPase in brown adipose tissue is the result of post‐transcriptional regulation

Temperature Threshold and Preservation of Signaling for Mitochondrial Membrane Proteins during Ischemia in Rabbit Heart

Mitochondrial ATP synthase F1‐β‐subunit is a calcium‐binding protein

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Mitochondrial ATP synthase F₁‐β‐subunit is a calcium‐binding protein