Structural and Kinetic Characterization of Quinolinate Phosphoribosyltransferase (hQPRTase) from Homo sapiens

Human quinolinate phosphoribosyltransferase (EC 2.4.2.19) (hQPRTase) is a member of the type II phosphoribosyltransferase family involved in the catabolism of quinolinic acid (QA). It catalyses the formation of nicotinic acid mononucleotide from quinolinic acid, which involves a phosphoribosyl trans...

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Bibliographic Details
Published in:Journal of molecular biology 2007-10, Vol.373 (3), p.755-763
Main Authors: Liu, Huanting, Woznica, Kerry, Catton, Gemma, Crawford, Amanda, Botting, Nigel, Naismith, James H.
Format: Article
Language:English
Subjects:
5-phosphoribosylpyrophosphate
Amino Acid Sequence
Binding Sites
Catalysis
Crystallography, X-Ray
Escherichia coli
Homo sapiens quinolinate phosphoribosyltransferase
Humans
Kinetics
Models, Molecular
Molecular Sequence Data
Mutagenesis, Site-Directed
NAD biosynthesis
nicotinic acid mononucleotide
Pentosyltransferases - chemistry
Pentosyltransferases - genetics
Pentosyltransferases - metabolism
Phosphoribosyl Pyrophosphate - metabolism
Protein Conformation
quinolinic acid
Sequence Homology, Amino Acid
Substrate Specificity
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Summary:Human quinolinate phosphoribosyltransferase (EC 2.4.2.19) (hQPRTase) is a member of the type II phosphoribosyltransferase family involved in the catabolism of quinolinic acid (QA). It catalyses the formation of nicotinic acid mononucleotide from quinolinic acid, which involves a phosphoribosyl transfer reaction followed by decarboxylation. hQPRTase has been implicated in a number of neurological conditions and in order to study it further, we have carried out structural and kinetic studies on recombinant hQPRTase. The structure of the fully active enzyme overexpressed in Escherichia coli was solved using multiwavelength methods to a resolution of 2.0 Å. hQPRTase has a α/β barrel fold sharing a similar overall structure with the bacterial QPRTases. The active site of hQPRTase is located at an α/β open sandwich structure that serves as a cup for the α/β barrel of the adjacent subunit with a QA binding site consisting of three arginine residues (R102, R138 and R161) and two lysine residues (K139 and K171). Mutation of these residues affected substrate binding or abolished the enzymatic activity. The kinetics of the human enzyme are different to the bacterial enzymes studied, hQPRTase is inhibited competitively and non-competitively by one of its substrates, 5-phosphoribosylpyrophosphate (PRPP). The human enzyme adopts a hexameric arrangement, which places the active sites in close proximity to each other.
ISSN:0022-2836
1089-8638
DOI:10.1016/j.jmb.2007.08.043