Drosophila melanogaster aldolase: Characterization of the isozymes α, β, and γ generated from a single gene

Rong Zhang, Tatsuo Kai, Yasushi Sugimoto, Takahiro Kusakabe, Yozo Takasaki, Katsumi Koga, Katsuji Hori

Research output: Contribution to journalArticle

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Abstract

Three isozymic forms, α, β, and γ, of Drosophila melanogaster aldolase are produced from a single gene by alternative usage of the triple exons 4 (4α, 4β, and 4γ) (Shaw-Lee et al. (1992) J.Biol. Chem. 267, 3959-3967; Kim et al. (1992) Mol. Cell. Biol. 12, 773-783; Kai et al. (1992) J. Biochem.112, 677-688). The expression plasmids for the respective isozymes were transfected into Escherichia coli cells, and the isozymes α and β were purified to homogeneity by a simple procedure, though isozyme γ was only partially purified. These isozymes are active towards two substrates, fructose-1, 6-bisphosphate (Fru-1, 6-P2) and fructose-1-phosphate (Fru-1-P), with a preference for Fru-1,6-P2 over Fru-1-P, but they have different kcat/Km values towards these two substrates; isozyme α shows the highest kcat/Km value for Fru-1-P among the three isozymes, whereas isozyme β has the highest value for Fru-1,6-P2. These isozymes show similarity in optimal pHs, thermalstability, and Km values for both Fru-1,6-P2 and Fru-1-P. They are composed of four identical subunits of 40 kDa, forming a tetramer with a molecular weight of approximately 160 kDa. The three isozymes are different in primary structure only at the carboxyl-terminal region encoded by the respective exon 4. Therefore, this region should be primarily responsible for the distinct characteristics of these isozymes.

Original languageEnglish
Pages (from-to)183-188
Number of pages6
JournalJournal of biochemistry
Volume118
Issue number1
DOIs
Publication statusPublished - Jan 1 1995
Externally publishedYes

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Fructose-Bisphosphate Aldolase
Drosophila melanogaster
Isoenzymes
Genes
Exons
Substrates
Escherichia coli
Plasmids
Molecular Weight
Molecular weight

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Cite this

Drosophila melanogaster aldolase : Characterization of the isozymes α, β, and γ generated from a single gene. / Zhang, Rong; Kai, Tatsuo; Sugimoto, Yasushi; Kusakabe, Takahiro; Takasaki, Yozo; Koga, Katsumi; Hori, Katsuji.

In: Journal of biochemistry, Vol. 118, No. 1, 01.01.1995, p. 183-188.

Research output: Contribution to journalArticle

Zhang, Rong ; Kai, Tatsuo ; Sugimoto, Yasushi ; Kusakabe, Takahiro ; Takasaki, Yozo ; Koga, Katsumi ; Hori, Katsuji. / Drosophila melanogaster aldolase : Characterization of the isozymes α, β, and γ generated from a single gene. In: Journal of biochemistry. 1995 ; Vol. 118, No. 1. pp. 183-188.
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