Enzyme Activity Measurement of Hypoxanthine Phosphoribosyltransferase Using Spectrophotometric Assays

Creative Enzymes is distinguished by its extraordinary techniques and expertise. Powered by the cutting-edge equipment, Creative Enzymes can provide excellence assay services for transferases. We have deep understandings of spectrophotometric assays and are excited to offer accurate and reliable assay services for hypoxanthine phosphoribosyltransferase.

Hypoxanthine phosphoribosyltransferase (HPRT) has a wide range of distribution among organisms. It catalyzes the magnesium-dependent reversible transfer of a phosphoribosyl group from 5-phospho-α-D-ribosyl-1-pyrophosphate (PRPP) to a purine base to form a purine ribonucleotide, inosine monophosphate (IMP), or guanosine monophosphate (GMP).

As commonly known, purine nucleotides are essential for synthesis of DNA and RNA. In addition, they provide energy and serve as cofactors. Two pathways are known for the biosynthesis of purine nucleotides: the de novo and the salvage pathways. The salvage pathway benefits from the lower energy cost compared with the de novo synthesis of nitrogen bases. Hypoxanthine phosphoribosyltransferase (HPRT, EC 2.4.2.8) is a key enzyme in the purine salvage pathway. Due to high deficiency of the de novo pathway, many parasite organisms rely on HPRT to salvage exogenous purine bases to synthesize purine nucleotides. Therefore, this enzyme is critical to the survival of parasite protozoans, so that it has been proposed as a potential target for designing therapeutic inhibitors against parasite. As for humans, HPRT is not essential but a genetic defect of its activity has significant impacts on health. The complete deficiency of HPRT is associated with an X-chromosome-linked disorder, the Lesch-Nyhan syndrome, which is characterized clinically by hyperuricemia, mental retardation, choreoathetosis, and compulsive self-mutilation. A partial deficiency of HPRT is also associated with hyperuricemia, which results in the development of gout and nephrolithiasis. Therefore, more and more studies are performed aimed at understanding the catalytic mechanism, which is related to clinical diseases. The accurate activity assay methods will make great contributions to the research, and Creative Enzymes can support such research with reliable assay services by spectrophotometric analysis.

Figure: The crystal structure of HPRT from Bacillus anthracis. PDB: 3O7M

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