Enzyme Activity Measurement of Adenine Phosphoribosyltransferase Using Chromatographic Assays

Creative Enzymes has been active in the enzyme industry for years. Through these years, we have accumulated extensive experiences and grasped various experimental skills. Our strong presence in enzyme testing guarantees reliable methods and precise results. Based on the up-to-date equipment, Creative enzymes provides the top services for adenine phosphoribosyltransferase by chromatographic assays.

Adenine phosphoribosyltransferase (APRT) is a member of the phosphoribosyltransferase (PRT) family. The responsibility of the enzyme is recycling the purine base adenine by catalyzing the Mg2+-dependent conversion of adenine and α-D-5-phosphoribosyl-1-pyrophosphate (PRPP) to adenosine monophosphate (AMP) and pyrophosphate.

In mammals, APRT is present in all tissues and involved in the metabolic salvage of adenine resulting from the polyamine biosynthesis pathway or from dietary sources. In organisms that can synthesize purines de novo, recycling of purines is primarily important during the period of rapid growth. An example is rapidly dividing cancer cells. Although APRT is not essential for humans, a genetic defect of its activity is not without consequence. Two types of APRT deficiencies are currently recognized in humans, both due to mutations in the corresponding gene. Type I deficiency is the result of a complete loss of APRT activity, whereas type II deficiency APRT leads to a tenfold-reduced affinity for its substrate. The major clinical manifestations for APRT-deficiency are the appearance of kidney stones, crystalluria, hematuria, dysuria, and infections in the urinary tract. Such clinical profiles can result in severe kidney failure. Therefore, APRT has become a judicious target for chemotherapeutic intervention. Considering the great significance of its clinical positions, an increasing number of studies have carried out on this enzyme. However, the activity measurement was hardly performed in the past because of the lack of suitable methods and precise detecting instrument. With the development of analytical techniques, the establishment of standard approaches has made great progresses. Creative Enzymes closely follows the updated skills and levels up our abilities to provide top services for APRT.

Figure: The crystal structure of human adenine phosphoribosyltransferase complexed with PRPP, ADE and R5P.

Creative Enzymes makes every effort toward the exploration and exploitation of enzymology services. Our particular abilities in activity testing have been approved by many customers. For enzyme activity quantification, Creative Enzymes will always be your most reliable partner.