Abstract
Ergothioneine is an N-α-trimethyl-2-thiohistidine derivative that occurs in human, plant, fungal, and bacterial cells. Biosynthesis of this redox-active betaine starts with trimethylation of the α-amino group of histidine. The three consecutive methyl transfers are catalyzed by the S-adenosylmethionine-dependent methyltransferase EgtD. Three crystal structures of this enzyme in the absence and in the presence of N-α-dimethylhistidine and S-adenosylhomocysteine implicate a preorganized array of hydrophilic interactions as the determinants for substrate specificity and apparent processivity. We identified two active site mutations that change the substrate specificity of EgtD 10(7)-fold and transform the histidine-methyltransferase into a proficient tryptophan-methyltransferase. Finally, a genomic search for EgtD homologues in fungal genomes revealed tyrosine and tryptophan trimethylation activity as a frequent trait in ascomycetous and basidomycetous fungi.
Keywords:
amino acids; betaines; biosynthesis; ergothioneine; hypaphorine; methyltransferases.
© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Ascomycota / enzymology*
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Ascomycota / genetics
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Basidiomycota / enzymology*
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Basidiomycota / genetics
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Betaine / chemistry*
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Betaine / metabolism
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Catalytic Domain
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Crystallography, X-Ray
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Ergothioneine / chemistry*
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Ergothioneine / metabolism
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Escherichia coli / genetics
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Escherichia coli / metabolism
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Fungal Proteins / chemistry*
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Fungal Proteins / genetics
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Fungal Proteins / metabolism
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Gene Expression
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Histidine / chemistry
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Histidine / metabolism
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Methyltransferases / chemistry*
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Methyltransferases / genetics
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Methyltransferases / metabolism
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Models, Molecular
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Molecular Sequence Data
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Mutation
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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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S-Adenosylhomocysteine / chemistry
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S-Adenosylhomocysteine / metabolism
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S-Adenosylmethionine / chemistry
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S-Adenosylmethionine / metabolism
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Sequence Alignment
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Sequence Homology, Amino Acid
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Substrate Specificity
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Tryptophan / chemistry
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Tryptophan / metabolism
Substances
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Fungal Proteins
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Recombinant Proteins
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Betaine
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Histidine
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S-Adenosylmethionine
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Tryptophan
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S-Adenosylhomocysteine
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Ergothioneine
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Methyltransferases