Abstract
The TEF4 gene of the non-saccharomyces yeast Yarrowia lipolytica encodes an EF1Bgamma protein with structural similarity to the glutathione transferases (GSTs). This 1203bp gene was cloned, over-expressed in Escherichia coli, and the recombinant protein characterized. DNA sequencing of the cloned gene agreed with the recently completed Y. lipolytica genome and showed 100% identity to a previously reported 30-residue N-terminal sequence for a 110kDa Y. lipolytica GST, except that it encoded two additional N-terminal residues (N-Met-Ser-). The recombinant protein (subunit M(r) 52kDa) was found not to possess GST activity with 1-chloro-2,4-dinitrobenzene. Partial tryptic digestion released two fragments of M(r) 22 and 18kDa, which we interpret as N- and C-terminal domains. Homology modeling confirmed that the N-terminal domain of Y. lipolytica TEF4 encodes a GST-like protein.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Cloning, Molecular
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Gene Expression
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Glutathione Transferase / chemistry*
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Glutathione Transferase / genetics
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Glutathione Transferase / metabolism*
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Models, Biological
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Models, Molecular
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Molecular Weight
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Peptide Elongation Factor 1
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Peptide Elongation Factors / chemistry
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Peptide Elongation Factors / genetics*
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Peptide Elongation Factors / metabolism
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Protein Structure, Tertiary
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Recombinant Proteins / biosynthesis
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Recombinant Proteins / chemistry
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Recombinant Proteins / genetics
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Recombinant Proteins / metabolism
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Saccharomyces cerevisiae / enzymology*
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Saccharomyces cerevisiae / genetics
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Saccharomyces cerevisiae Proteins / chemistry
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Saccharomyces cerevisiae Proteins / genetics*
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Saccharomyces cerevisiae Proteins / metabolism
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Structural Homology, Protein*
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Yarrowia / enzymology*
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Yarrowia / genetics
Substances
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CAM1 protein, S cerevisiae
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Peptide Elongation Factor 1
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Peptide Elongation Factors
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Recombinant Proteins
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Saccharomyces cerevisiae Proteins
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Glutathione Transferase