Molecular basis of classic galactosemia from the structure of human galactose 1-phosphate uridylyltransferase.

TitleMolecular basis of classic galactosemia from the structure of human galactose 1-phosphate uridylyltransferase.
Publication TypeJournal Article
Year of Publication2016
AuthorsMcCorvie, TJ, Kopec, J, Pey, AL, Fitzpatrick, F, Patel, D, Chalk, R, Shrestha, L, Yue, WW
JournalHum Mol Genet
Date Published2016 Jun 01
KeywordsBinding Sites, Catalytic Domain, Crystallography, X-Ray, Galactose, Galactosemias, Humans, Kinetics, Models, Molecular, Mutagenesis, Site-Directed, Mutation, Protein Conformation, Structure-Activity Relationship, Ternary Complex Factors, UTP-Hexose-1-Phosphate Uridylyltransferase

Classic galactosemia is a potentially lethal disease caused by the dysfunction of galactose 1-phosphate uridylyltransferase (GALT). Over 300 disease-associated GALT mutations have been reported, with the majority being missense changes, although a better understanding of their underlying molecular effects has been hindered by the lack of structural information for the human enzyme. Here, we present the 1.9 Å resolution crystal structure of human GALT (hGALT) ternary complex, revealing a homodimer arrangement that contains a covalent uridylylated intermediate and glucose-1-phosphate in the active site, as well as a structural zinc-binding site, per monomer. hGALT reveals significant structural differences from bacterial GALT homologues in metal ligation and dimer interactions, and therefore is a zbetter model for understanding the molecular consequences of disease mutations. Both uridylylation and zinc binding influence the stability and aggregation tendency of hGALT. This has implications for disease-associated variants where p.Gln188Arg, the most commonly detected, increases the rate of aggregation in the absence of zinc likely due to its reduced ability to form the uridylylated intermediate. As such our structure serves as a template in the future design of pharmacological chaperone therapies and opens new concepts about the roles of metal binding and activity in protein misfolding by disease-associated mutants.

Alternate JournalHum. Mol. Genet.
Citation Key10.1093/hmg/ddw091
PubMed ID27005423
PubMed Central IDPMC5081055