SANTI NURBAITI
Kelompok Keahlian Biokimia, Fakultas Matematika dan Ilmu Alam ITB.

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Journal : Jurnal Riset Kimia

SINTESIS DAN KARAKTERISASI NUKLEOTIDA BERTANDA [α-32P]ATP Rahman, Wira Yusril; Sarmini, Endang; Herlina, H.; Abidin, A.; Triyanto, T.; Hambali, H.; Nurbaiti, Santi
Jurnal Riset Kimia Vol 10, No 1 (2019): March
Publisher : Universitas Andalas

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.25077/jrk.v12i2.319

Abstract

The utilization of nuclear technology in health sector with molecular techniques is increasingly developed today, especially in Indonesia.  One of which is nucleotide compound marked with [α-32P]ATP, this compound has been used as tracer for deoxyribonucleic acid (DNA)/ ribonucleic acid (RNA) in the study of various physiological and pathological processes. [α-32P]ATP is synthesized through several stages of continuous reaction in one reaction vessel. It begins with synthesis of [γ-32P]ATP through an enzymatic reaction, using H332PO4 and ADP, and enzymes  of lactate dehydrogenase, 3-phosphoglycerate phosphokinase and glyceraldehide 3-phospho  dehydrogenase; followed by phosphorilation of 3’AMP with T4 polinucleotide kinase enzyme to produce 3’-[5’-32P]ATP. The result is hydrolyzed with nuclease P1 enzyme to produce [5’-32P]AMP. The unreacted [γ-32P] is degraded by the addition of hexokinase enzyme and glucose. At the final stage of the reaction, the [5’-32P]AMP is  phosphorilated using phosphoenol-piruvat, piruvat kinase, and myokinase to produce [α-32P]ATP. The test results show that the every stage of reaction is characterized using TLC method, PEI cellulose paper as stationary phase and KH3PO4 0,5 M pH 3,5 as mobile phase. At the end of reaction, the yield of [α-32P]ATP reaches 71,7%, at Rf = 0,2.