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Hybrid isozymes of rat pyruvate kinase. Their subunit structure and developmental changes in the liver 总被引:1,自引:0,他引:1
Thin-layer polyacrylamide gel electrophoresis of various rat tissues revealed three major isozymes (types L, M1 and M2) and various intermediate forms of pyruvate kinase (ATP: pyruvate 2-O-phosphotransferase, EC 2.7.1.40). In vitro dissociation and reassociation of purified enzymes showed that the three major isozymes had homotetrameric structures. L.M2 hybrids and M1.M2 hybrids closely resembled some naturally occurring intermediates; the subunit structure of intermediates isolated from the small intestine (form 3 or form 4) were estimated to be (L)2(M2)2 and (L)(M2)3, respectively. Pyruvate kinase activity after electrophoresis could be estimated quantitatively from densitometric measurements of the electrophoretic pattern. Type L activity in fetal liver was separated from type R activity derived from intrahepatic erythropoietic cells. It changes in three distinct steps during development: it increased during the late fetal period, remained steady during the neonatal period and increased again after weaning. Some of the intermediates found in extracts of early fetal iver were shown to cross-react with both anti-L and anti-M1 serum, suggesting that they might be L.M2 or R.M2 hybrids. These hybrid enzymes were shown to appear only during early fetal and neonatal periods. 相似文献
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The relative rates of synthesis and degradation for liver pyruvate kinase have been determined in rats fed standard lab chow, fasted, and refed a high-carbohydrate-low-protein diet. Relative rates of synthesis and apparent rates of degradation were determined by pulse-labeling the enzyme in vivo with l-[4,5-3H]leucine and by measuring the incorporation of radioactivity into liver pyruvate kinase after quantitative precipitation of the enzyme with anti-liver pyruvate kinase immunoglobulin. The relative rate of synthesis decreased approximately 75% upon fasting and then increased 20- to 30-fold upon refeeding the high-carbohydrate diet. The apparent half-lives for liver pyruvate kinase in fasted, control, and refed animals are very similar (55, 59, and 47 h, respectively). Thus, the nutritional alterations in the levels of liver pyruvate kinase seem to result primarily from alterations in the rate of enzyme synthesis. 相似文献
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H Muirhead D A Clayden D Barford C G Lorimer L A Fothergill-Gilmore E Schiltz W Schmitt 《The EMBO journal》1986,5(3):475-481
The complete amino acid sequence of cat muscle pyruvate kinase has been determined and fitted to the 2.6 A resolution electron density map. Residues in the active site region are highly conserved in the cat muscle, chicken muscle, rat liver and yeast enzymes. The enzyme-bound magnesium, which is essential for activity, interacts with the side chain of glutamate-271 and with two main carbonyl groups. Lysine-269 is the probable acid/base catalyst responsible for the interconversion of pyruvate and enolpyruvate. A possible binding site for the essential monovalent cation is proposed. 相似文献
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Rat liver pyruvate kinase, prepared by Sephadex treatment of a 10(5)g supernatant in phosphate buffer, is quite stable and gives reproducible results when a variety of parameters are altered in the enzyme assay. Incubation of this preparation at 25 degrees C or 0-2 degrees C has no effect on the activation by fructose 1,6-diphosphate or inhibition by ATP or alanine. 相似文献
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Bror Edlund Jill Andersson Vincent Titanji Ulla Dahlqvist Pia Ekman Örjan Zetterqvist Lorentz Engström 《Biochemical and biophysical research communications》1975,67(4):1516-1521
One dominating peptic phosphopeptide, Asx-Thr-Lys-Gly-Pro-Glx-Ile-Glx-Thr-Gly-Val-Leu-Arg-Arg-Ala-(32P)SerP-Val-Ala-Glx-Leu, was obtained from rat liver pyruvate kinase (type L) phosphorylated by cyclic 3′,5′-AMP-stimulated protein kinase from the same tissue. The sequence around the phosphorylated serine residue is similar to that of a corresponding but smaller peptic phosphopeptide previously isolated from pig liver (type L) pyruvate kinase, Leu-Arg-Arg-Ala-(32P)SerP-Leu. 相似文献
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After 5 h of treatment with glucagon, liver L-type pyruvate kinase (ATP: pyruvate 2-0-phosphotransferase; EC 2.7.1.40) showed a significant decrease of K0.5 and the Hill coefficient (nH) in the absence of fructose 1,6-diphosphate. However, in the presence of fructose 1,6-diphosphate, liver enzymes from treated rats showed a slight decrease of K0.5 but nH remained unchanged. In both circumstances, no changes of Vmax were observed after treatment. These changes in the kinetic properties of liver L-type pyruvate kinase are consistent with the dephosphorylation of the enzyme caused by insulin release in response to treatment with glucagon. 相似文献
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D. J. H. Brock 《Human genetics》1970,10(1):30-34
Summary A modified technique has been introduced which allows detection of PK in the presence of AK after starch-gel electrophoresis. It is shown that the PK isoenzyme patterns of different human tissues are independent of the AK phenotype of the person concerned. It is concluded that there is no basis for the previously held assumption that these two enzymes share a common polypeptide chain.This work was supported by a grant from the Scottish Hospital Endowments Research Trust (HERT 309). 相似文献
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Mark L. Johnson Joell B. Hansen James C. Donofrio Carlo M. Veneziale 《Biochimica et Biophysica Acta (BBA)/General Subjects》1981,675(1):140-142
Dexamethasone in the medium perfusin isolated rabbit livers caused a fast-acting and reversible effect on liver pyruvate kinase. The effect was to lower th assayable V activity (units/g tissue) without changing the concentration (nmol/g enzyme protein). In effect, glucocorticoid lowered the specific activity (units/nmol of enzyme) by direct action on liver. The effect on liver pyruvate kinase is mediated by a relatively stable alteration; 30 min after perfusate (with steroid) was replaced by perfusate (without steroid), the effect remained strongly evident. 相似文献
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The transport of pyruvate in rat liver mitochondria 总被引:10,自引:0,他引:10
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Yeast pyruvate kinase. Native and subunit molecular weight 总被引:2,自引:0,他引:2
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The ontogenetic study of pyruvate kinase in the brain and liver tissues was performed in different batches of rats, from the fœtus at the 13th day of gestation to the adult subject.
- — According to the kinetic study, the shape of the curve is transformed from sigmoid to hyperbolic from the 13th day of fœtal life to adulthood in the brain. Hill cœfficient increases with the age of animal in the liver tissue.
- — According to polyacrylamide gel isoelectrofocusing, a family of four, transitory or definite bands are detected in the fœtal brain: they are well defined by their pHi: M4, M3, M2, M1; at the adult stage, M1 predominates, M2 is minor. Three principal bands are distinguished in the liver: two are characteristic of fœtal life (Lf and M2), one of adulthood (L).
- — According to the immunochemical tests, there are antigenic determinants common to M1, M2, M3 and M4.
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Evidence for the presence of two types of pyruvate kinase in rat liver 总被引:13,自引:0,他引:13
T Tanaka Y Harano H Morimura R Mori 《Biochemical and biophysical research communications》1965,21(1):55-60
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