Abstract
Limited proteolysis of the arom enzyme complex of Neurospora crassa by trypsin or subtilisin yielded a stable fragment of Mr 68000. This fragment, which was purified by two-dimensional polyacrylamide-gel electrophoresis, was shown by activity staining to contain the shikimate dehydrogenase active site, and by substrate labelling with 3-dehydroquinate and NaB3H4 to contain the 3-dehydroquinase active site. The fragment thus constitutes a bifunctional domain containing the two enzymic activities that are known, from genetic evidence, to be located adjacently at the C-terminal end of the pentafunctional arom polypeptide.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Alcohol Oxidoreductases*
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Binding Sites
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Chromatography, Affinity
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Electrophoresis, Polyacrylamide Gel
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Hydro-Lyases / isolation & purification
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Ligands
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Lyases*
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Models, Chemical
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Multienzyme Complexes / antagonists & inhibitors
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Multienzyme Complexes / isolation & purification*
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Neurospora / enzymology*
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Neurospora crassa / enzymology*
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Peptides / analysis
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Phosphotransferases (Alcohol Group Acceptor)*
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Subtilisins
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Transferases*
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Trypsin
Substances
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Ligands
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Multienzyme Complexes
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Peptides
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arom enzyme
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Alcohol Oxidoreductases
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Transferases
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Phosphotransferases (Alcohol Group Acceptor)
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Subtilisins
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Trypsin
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Lyases
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Hydro-Lyases
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3-dehydroquinate dehydratase