A modified method for synthesis of [γ-32P] labelled adenosine triphosphate
- 106 Downloads
Abstract
Production of [γ-32P]-ATP using three glycolysis enzymatic reaction i.e. glyceraldehyde 3-phosphate dehydrogenase, 3-phosphoglyceric phosphokinase and lactate dehydrogenase has been conducted. dl-glyceraldehyde 3-phosphate, Adenosine Diphosphate and H 3 32 PO4 was used as precursors for this reaction. Purification of [γ-32P]-ATP was performed by using DEAE-Sephadex column chromatography. The result suggested that this simple method could be used for producing [γ-32P]-ATP to support the provision of radiolabeled nucleotide for biotechnology research in Indonesia.
Keywords
Labeled nucleotide [γ-32P]-ATP Synthesis Enzymatic reaction DEAE-Sephadex column chromatographyNotes
Acknowledgments
The author wish to thank to Dra. Siti Darwati, Drs. Hotman Lubis and to all those who have given advice, motivation and support both materially and spiritually, so the author can finish writing scientific papers.
References
- 1.Lehninger AL (1982) Principles of Biochemistry. Worth Publisher, Inc, New YorkGoogle Scholar
- 2.Syaifudin M, Tetriana D (2007), Analysis of mutation of inhA gene for examining the resistance of M. tuberculosis to isoniazid with radioactive SSCP method, CTRSM - BATAN, Jakarta, 1–9Google Scholar
- 3.Maria Lina R, Bela B, Yasmon A (2009) KatG gene mutation detection (MyobacteriumTuberculosis) by PCR method (Polymerase Chain Reaction)—dot blot hybridization using 32P labelled oligonucleotides tracers. J Ilmiah AI & R 5(1):174–180Google Scholar
- 4.Budiawan (2000), Development of a 32P-postlabelling technique to detect early cancer risk, proceedings of scientific meeting of the research and development of isotopes and radiation technology, 39–44Google Scholar
- 5.Sakamoto F, Izumo M, Hashimoto K, Fujii Y (1998) Study of optimum condition for synthesis of [γ-32P]-ATP with high specific radioactivity. J Radioanal Nucl Chem 239(2):423–427CrossRefGoogle Scholar
- 6.Schendel Paul F, Wells Robert D (1973) The synthesis and purification of [γ-32P] adenosine triphosphate with high specific activity. J Biol Chem 248(23):8319–8321Google Scholar
- 7.Jhonson RA, Walseth TA (1980) “Enzymatic process for preparing [γ-32P]-labeled nucleotides”, United State Patent, JuneGoogle Scholar
- 8.Rahman WY, Sarmini E, Herlina, Triyanto, Mutalib A, Nurbaiti S (2011) Synthesis of 32P-labeled ATP for molecular biology tracer. J Radioiosot Radiopharm 14(1):9–16Google Scholar
- 9.Madden Mark, Song Christine, Tse Kathy, Wong Ambrose (2004) The inhibitory effect of EDTA and Mg2+ on the activity of nadh dehydrogenase in lysozyme lysis. JEMI 5:8–15Google Scholar