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Bovine hexokinase type I: Full-length cDNA sequence and characterisation of the recombinant enzyme

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Abstract

This study reports the revised and full-length cDNA sequence of bovine hexokinase type I obtained from bovine brain. Since dissimilarities have been observed between the published bovine hexokinase type I coding sequence (GenBank accession no. M65140) (Genomics 11: 1014-1024, 1991) and an analysed portion of bovine hexokinase type I gene, the entire open reading frame was re-sequenced and the ends of cDNA isolated by rapid amplification of cDNA ends. The coding sequences, when compared with the published bovine hexokinase type I, contained a large number of mismatches that lead to changes in the resulting amino acid sequence. The revisions result in a hexokinase type I cDNA of 3619 bp that encodes a protein of 917 amino acids highly homologous to human hexokinase type I. The expression of the recombinant full-length enzyme demonstrated that it was a catalytically active hexokinase. When characterised for its kinetic and regulatory properties, it displayed the same affinity for glucose and MgATP as the human hexokinase type I and was inhibited by glucose 6-phosphate competitively versus MgATP. The production of the N- and C-terminal recombinant halves of the enzyme followed by comparison with the full-length hexokinase indicated that the catalytic activity is located in the C-terminal domain. (Mol Cell Biochem 268: 9–18, 2005)

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Authors and Affiliations

  1. Centro di Biotecnologie, University of Urbino, via Campanella, 1, Fano, Italy

    Francesca Andreoni & Mauro Magnani Prof.

  2. Istituto di Chimica Biologica “G. Fornaini, University of Urbino, via Saffi, 2, Urbino, Italy

    Giordano Serafini, Maria Elena Laguardia & Mauro Magnani Prof.

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  1. Francesca Andreoni
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  2. Giordano Serafini
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  3. Maria Elena Laguardia
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  4. Mauro Magnani Prof.
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Correspondence to Mauro Magnani Prof..

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Andreoni, F., Serafini, G., Laguardia, M.E. et al. Bovine hexokinase type I: Full-length cDNA sequence and characterisation of the recombinant enzyme. Mol Cell Biochem 268, 9–18 (2005). https://doi.org/10.1007/s11010-005-1846-5

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  • DOI: https://doi.org/10.1007/s11010-005-1846-5

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