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PKU mutation p.G46S prevents the stereospecific binding of l-phenylalanine to the dimer of human phenylalanine hydroxylase regulatory domain.


ABSTRACT: Mammalian phenylalanine hydroxylase (PAH) has a potential allosteric regulatory binding site for l-phenylalanine (l-Phe), in addition to its catalytic site. This arrangement is supported by a crystal structure of a homodimeric truncated form of the regulatory domain of human PAH (hPAH-RD 1-118/19-118) [Patel D et al. (2016) Sci Rep doi: 10.1038/srep23748]. In this study, a fusion protein of the domain (MBP-(pepXa)-hPAH-RD 1-120) was overexpressed and recovered in a metastable and soluble state, which allowed the isolation of a dimeric and a monomeric fusion protein. When cleaved from MBP, hPAH-RD forms aggregates which are stereospecifically inhibited by l-Phe (> 95%) at low physiological concentrations. Aggregation of the cleaved dimer of the mutant form hPAH-G46S-RD was not inhibited by l-Phe, which is compatible with structurally/conformationally changed ?????? ACT domain folds in the mutant.

SUBMITTER: Leandro J 

PROVIDER: S-EPMC5292662 | biostudies-literature | 2017 Feb

REPOSITORIES: biostudies-literature

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PKU mutation p.G46S prevents the stereospecific binding of l-phenylalanine to the dimer of human phenylalanine hydroxylase regulatory domain.

Leandro João J   Saraste Jaakko J   Leandro Paula P   Flatmark Torgeir T  

FEBS open bio 20170121 2


Mammalian phenylalanine hydroxylase (PAH) has a potential allosteric regulatory binding site for l-phenylalanine (l-Phe), in addition to its catalytic site. This arrangement is supported by a crystal structure of a homodimeric truncated form of the regulatory domain of human PAH (hPAH-RD <sup>1-118/19-118</sup>) [Patel D <i>et al</i>. (2016) <i>Sci Rep</i> doi: 10.1038/srep23748]. In this study, a fusion protein of the domain (MBP-(pep<sub>Xa</sub>)-hPAH-RD <sup>1-120</sup>) was overexpressed an  ...[more]

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