Abstract
Phenylalanine hydroxylase (PAH) is activated by its substrate phenylalanine and inhibited by its cofactor tetrahydrobiopterin (BH(4)). The crystal structure of PAH revealed that the N-terminal sequence of the enzyme (residues 19-29) partially covered the enzyme active site, and suggested its involvement in regulation. We show that the protein lacking this N-terminal sequence does not require activation by phenylalanine, shows an altered structural response to phenylalanine, and is not inhibited by BH(4). Our data support the model where the N-terminal sequence of PAH acts as an intrasteric autoregulatory sequence, responsible for transmitting the effect of phenylalanine activation to the active site.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Binding Sites
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Biopterins / analogs & derivatives*
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Biopterins / metabolism
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Biopterins / pharmacology
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Chymotrypsin / metabolism
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Enzyme Activation / drug effects
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Models, Molecular
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Phenylalanine / antagonists & inhibitors
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Phenylalanine / metabolism
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Phenylalanine / pharmacology
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Phenylalanine Hydroxylase / antagonists & inhibitors
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Phenylalanine Hydroxylase / chemistry*
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Phenylalanine Hydroxylase / genetics
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Phenylalanine Hydroxylase / metabolism*
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Phosphorylation / drug effects
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Protein Conformation
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Recombinant Proteins / antagonists & inhibitors
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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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Sequence Deletion / genetics
Substances
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Recombinant Proteins
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Biopterins
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Phenylalanine
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Phenylalanine Hydroxylase
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Chymotrypsin
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sapropterin