Human liver type pyruvate kinase: complete amino acid sequence and the expression in mammalian cells
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TLDR
The full-length sequence of human L-type PK cDNA was isolated and determined and activity was detected in the extract of COS cells by the classical PK electrophoresis method.Abstract:
Pyruvate kinase (PK) has four isozymes (L, R, M1, M2) that are encoded by two different genes. Among these isozymes, abnormalities of liver (L)-type PK is considered to be associated with hereditary nonspherocytic hemolytic anemia in humans. We isolated and determined the full-length sequence of human L-type PK cDNA. The cDNA contains 1629 base pairs encoding 543 amino acids, 68 base pairs of 5'-noncoding sequence, and 734 base pairs of 3'-noncoding sequence. The similarity between human and rat L-type PK was 86.9% at the nucleotide sequence level and 92.4% at the amino acid sequence level. The full-length L-type PK cDNA was placed under the promoter of simian virus 40 and introduced into monkey COS cells. Human L-type PK activity was detected in the extract of COS cells by the classical PK electrophoresis method.read more
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Journal ArticleDOI
The energy-less red blood cell is lost: erythrocyte enzyme abnormalities of glycolysis
TL;DR: This review summarizes the current knowledge regarding the genetic, biochemical, and structural features of clinically relevant red blood cell enzymopathies involved in the Embden-Meyerhof pathway and the Rapoport-Luebering shunt.
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Red cell pyruvate kinase deficiency: molecular and clinical aspects
TL;DR: The comparison of the recombinant mutants of human red cell PK with the wild‐type enzyme has enabled the effects of amino acid replacements on the enzyme molecular properties to be determined and help to correlate genotype to clinical phenotype.
Journal ArticleDOI
Pyruvate kinase: current status of regulatory and functional properties.
TL;DR: The results of a current and extensive review that determines the sites of conservation and/or difference in PK sequences, and the differences in the functional and regulatory properties of the enzymes are described.
Journal ArticleDOI
Pyruvate kinase deficiency: the genotype-phenotype association.
TL;DR: The clinical manifestations of red cell enzyme defects are not merely dependent on the molecular properties of the mutant protein but rather reflect the complex interactions of additional factors, including genetic background, concomitant functional polymorphisms of other enzymes, posttranslational or epigenetic modifications, ineffective erythropoiesis and differences in splenic function.
Journal ArticleDOI
Red cell pyruvate kinase deficiency: from genetics to clinical manifestations
Alberto Zanella,Paola Bianchi +1 more
TL;DR: Further data on clinical features of homozygous patients are needed, at least for some mutations, to allow a more precise genotype/phenotype correlation and analysis of the three-dimensional structure of the enzyme may help in predicting the severity of the molecular defect.
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Molecular Cloning: A Laboratory Manual
TL;DR: Molecular Cloning has served as the foundation of technical expertise in labs worldwide for 30 years as mentioned in this paper and has been so popular, or so influential, that no other manual has been more widely used and influential.
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Human liver type pyruvate kinase: cDNA cloning and chromosomal assignment.
Kenzaburo Tani,Fujii H,Hisashi Tsutsumi,Jun Sukegawa,Kumao Toyoshima,Michihiro C. Yoshida,Tamio Noguchi,Takehiko Tanaka,Shiro Miwa +8 more
TL;DR: Comparisons of the sequence data with the rat PK cDNAs indicated that the cDNA encoded information for the carboxyl terminal 105 amino acids of a human L-type PK and a 3' untranslated region of 734 nucleotides.