SINTESIS DAN KARAKTERISASI NUKLEOTIDA BERTANDA [α-32P]ATP

Authors

  • Wira Yusril Rahman Center for Radioisotope and Radiopharmaceuticals Technology (PTRR) National Nuclear Energy Agency of Indonesia (BATAN), Indonesia
  • Endang Sarmini Center for Radioisotope and Radiopharmaceuticals Technology (PTRR) National Nuclear Energy Agency of Indonesia (BATAN), Indonesia
  • H. Herlina Center for Radioisotope and Radiopharmaceuticals Technology (PTRR) National Nuclear Energy Agency of Indonesia (BATAN), Indonesia
  • A. Abidin Center for Radioisotope and Radiopharmaceuticals Technology (PTRR) National Nuclear Energy Agency of Indonesia (BATAN), Indonesia
  • T. Triyanto Center for Radioisotope and Radiopharmaceuticals Technology (PTRR) National Nuclear Energy Agency of Indonesia (BATAN), Indonesia
  • H. Hambali Center for Radioisotope and Radiopharmaceuticals Technology (PTRR) National Nuclear Energy Agency of Indonesia (BATAN), Indonesia
  • Santi Nurbaiti Chemistry Department Faculty of Mathematics and Natural Sciences Bandung Institute of Technology, Indonesia

DOI:

https://doi.org/10.25077/jrk.v12i2.319

Keywords:

tracer, labelled nucleotide[α-32P]ATP, synthesis, enzymatic reaction

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.

References

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Published

2019-04-14

How to Cite

Rahman, W. Y., Sarmini, E., Herlina, H., Abidin, A., Triyanto, T., Hambali, H., & Nurbaiti, S. (2019). SINTESIS DAN KARAKTERISASI NUKLEOTIDA BERTANDA [α-32P]ATP. Jurnal Riset Kimia, 10(1), 30–35. https://doi.org/10.25077/jrk.v12i2.319

Issue

Vol. 10 No. 1 (2019): March

Section

Articles

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