MODIFIKASI SINTESIS NUKLEOTIDA BERTANDA [γ-32 P] ATP

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ABSTRAK

MODIFIKASI SINTESIS NUKLEOTIDA BERTANDA [Y-32P]ATP.

Dalam perkembangan biologimolekul, radionuklida dalam bentuk senyawa bertanda telah digunakan sebagai perunut deoxyribonucleicacid (DNA)/ribonucleic acid (RNA) untuk mendalami berbagai macam proses fisiologi dan patologi. Salahsatu senyawa tersebut adalah nukleotida bertanda fosfor-32 (32P) [γ-32P]-adenosine triphosphate {[γ-32P]-ATP} yang banyak digunakan dalam penelitian biologi molekul. Untuk dapat menunjang penelitian biologimolekul di Indonesia, pada penelitian ini telah dilakukan pembuatan senyawa nukleotida bertanda [γ-32P]-ATP melalui reaksi enzimatis dengan melakukan modifikasi pada metoda sintesisnya menggunakanprekursor DL-glyceraldehyde 3-phosphate, nukleotida adenosine di-phosphate (ADP) dan H332PO4, sertaenzim gliseraldehid 3-phosphat dehidrogense, 3-phosphogliserat-kinase dan laktat dehidrogenase. Pemurnian[γ-32P]-ATP hasil sintesis dengan menggunakan kolom kromatografi DEAE-Sephadex. Dari proses sintesisdan pemurnian yang telah dilakukan berhasil diperoleh [γ-32P]-ATP dengan aktifitas 1,175 mCi dankemurnian radiokimia 99,49%. Dengan berhasilnya dilakukan sintesis dan pemurnian [γ-32P]-ATP, makaPusat Radiosiotop dan Radiofarmaka akan dapat menyediakan nukleotida bertanda dimaksud di atas untukmenunjang penelitian biologi molekul di Indonesia.

Kata Kunci: nukleotida bertanda [γ-32P]ATP, sintesis, reaksi enzimatis, pemurnianABSTRACTMODIFICATION OF SYNTHESIS NUCLEOTIDES [Y-32P] ATP.

In molecular biology, radionuclidesin the form of radiolabeled compounds have been widely used as deoxyribonucleic acid (DNA) / ribonucleicacid (RNA) tracer in order to explore a wide range of physiological and pathological processes. One of suchcompounds is [γ-32P]-adenosine triphosphate {[γ-32P]-ATP} [γ-32P]-ATP which has been widely used in thebiotechnology research. In order to support the biotechnology research in Indonesia in this project, [γ-32P]-ATP had been synthesized by enzymatic reactions with modifying the method of synthesis using theprecursor DL-glyceraldehydde 3-phosphate, nucleotides Adenosine Diphosphate (ADP) and H332PO4 andenzymes glyceraldehid 3-phosphate dehydrogenase, 3-phosphoglyceryc phosphokinase and lactatedehydrogenase. The purification of the synthesized [γ-32P]-ATP, by using DEAE Sephadex columnchromatography.  The synthesis and purification process that had been performed were able in producing of[γ-32P]-ATP with radioactivity of 1,175 mCi and  radiochemical purity of 99,49%.. Having successfullyprepared the [γ-32P]-ATP and application, in the near future the Radioiotopes and RadiopharmaceuticalsCentre is expected to be able in providing the above-mentioned radiolabeled nucleotide for biotechnologyresearch in Indonesia.

Key words : labeled nucleotide [γ-32P]-ATP, synthesis, enzimatic reaction, purification