UPRT Human

The human recombinant form of uracil phosphoribosyltransferase was identified and characterized through various studies. Initially, this enzyme was reported in plants like Arabidopsis thaliana, but not in mammals . However, a novel family of uracil phosphoribosyltransferase was later discovered in humans. The gene encoding this enzyme was isolated from a human fetal brain library and named human UPRTase .

The open reading frame (ORF) of the human UPRTase gene was cloned into the pQE30 vector and expressed in Escherichia coli M15 cells. The protein was then purified using Ni-NTA affinity chromatography . Despite successful purification, the enzymatic activity of UPRTase could not be detected through spectrophotometry .

Reverse transcription polymerase chain reaction (RT-PCR) analysis revealed that human UPRTase is expressed in various tissues, including the prostate, heart, brain, lung, and skeletal muscle . Subcellular localization studies using UPRTase-EGFP fusion protein showed that the enzyme is distributed in both the nucleus and cytoplasm of AD293 cells .

Evolutionary tree analyses indicated that UPRTase is conserved across different organisms. While UPRTase activity was observed in archaebacteria and eubacteria, higher organisms like Caenorhabditis elegans lacked two amino acids in the uracil-binding region, resulting in no UPRTase activity .

Human recombinant UPRTase has potential applications in the rational design of drugs for treating parasitic infections and cancer . The enzyme’s role in the pyrimidine salvage pathway makes it a target for developing antimetabolite drugs, which can inhibit nucleotide synthesis in rapidly dividing cells, such as cancer cells .