Files and links (2)
Published, Version of Record (VoR)CC BY V4.0, Open
Journal article
Biophysical characterization of full-length human phenylalanine hydroxylase provides a deeper understanding of its quaternary structure equilibrium
The Journal of biological chemistry, v 294(26), pp 10131-10145
28 Jun 2019
PMID: 31076506
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
Dysfunction of human phenylalanine hydroxylase (hPAH, EC 1.14.16.1) is the primary cause of phenylketonuria, the most common inborn error of amino acid metabolism. The dynamic domain rearrangements of this multimeric protein have thwarted structural study of the full-length form for decades, until now. In this study, a tractable C29S variant of hPAH (C29S) yielded a 3.06 Å resolution crystal structure of the tetrameric resting-state conformation. We used size-exclusion chromatography in line with small-angle X-ray scattering (SEC-SAXS) to analyze the full-length hPAH solution structure both in the presence and absence of Phe, which serves as both substrate and allosteric activators. Allosteric Phe binding favors accumulation of an activated PAH tetramer conformation, which is biophysically distinct in solution. Protein characterization with enzyme kinetics and intrinsic fluorescence revealed that the C29S variant and hPAH are otherwise equivalent in their response to Phe, further supported by their behavior on various chromatography resins and by analytical ultracentrifugation. Modeling of resting-state and activated forms of C29S against SAXS data with available structural data created and evaluated several new models for the transition between the architecturally distinct conformations of PAH and highlighted unique intra- and inter-subunit interactions. Three best-fitting alternative models all placed the allosteric Phe-binding module 8-10 Å farther from the tetramer center than do all previous models. The structural insights into allosteric activation of hPAH reported here may help inform ongoing efforts to treat phenylketonuria with novel therapeutic approaches.
Metrics
Details
- Title
- Biophysical characterization of full-length human phenylalanine hydroxylase provides a deeper understanding of its quaternary structure equilibrium
- Creators
- Emilia C Arturo - Temple University Health SystemKushol Gupta - University of PennsylvaniaMichael R Hansen - Temple University Health SystemElias Borne - Temple University Health SystemEileen K Jaffe - Temple University Health System
- Publication Details
- The Journal of biological chemistry, v 294(26), pp 10131-10145
- Publisher
- ASBMB Publications / Elsevier
- Grant note
- P30 CA006927 / NCI NIH HHS P30 GM133893 / NIGMS NIH HHS R01 NS100081 / NINDS NIH HHS
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Biochemistry and Molecular Biology
- Web of Science ID
- WOS:000474445600008
- Scopus ID
- 2-s2.0-85068652411
- Other Identifier
- 991019168224304721
UN Sustainable Development Goals (SDGs)
This publication has contributed to the advancement of the following goals:
InCites Highlights
Data related to this publication, from InCites Benchmarking & Analytics tool:
- Collaboration types
- Industry collaboration
- Domestic collaboration
- Web of Science research areas
- Biochemistry & Molecular Biology