Home > Journal > Efek Sitotoksik Suatu Protein Seperti Ribosome Inactivating Proteins yang Bersifat Asam dari Daun Mirabilis Jalapa L. Pada Sel Kanker

Efek Sitotoksik Suatu Protein Seperti Ribosome Inactivating Proteins yang Bersifat Asam dari Daun Mirabilis Jalapa L. Pada Sel Kanker

Cytotoxic effects of an acidic Ribosome-inactivating Protein like protein isolated from Mirabilis jalapa L. leaves on cancer cell-lines
Sudjadi *, Lucia D Witasari, Modesta T Sadarum, Nia Nastity dan Sismindari
Fakultas Farmasi Universitas Gadjah Mada Yogyakarta

Abstract
Most purification approaches of Ribosome-inactivating Proteins (RIPs) used a cation exchange chromatography resulting highly basic proteins, such as MJ-30 isolated from Mirabilis jalapa leaves. The M.jalapa unbounded protein fractions of the column showed cleavage activity on supercoiled double stranded DNA and possesed cytotoxic effects on cancer cell lines. So the purification and characterization of the acidic protein was needed to be done. Total proteins of M.jalapa leaves was collected from precipitating process of the extract using ammonium sulphate untill saturated. After dialysis, the proteins was applied into the weak base anion exchanger Ionenaustaucher Typ II. The bounded proteins was eluted with 0.0 to 0.5 M NaCl gradient in phosphate buffer. Active protein fractions were determined by supercoilded DNA cleavage activity and then cytotoxic activity was testes. Results showed that protein fraction eluted at 0.35 – 0.40 M NaCl possesed cleavage activity on supercoiled DNA. The acidic protein, MJ-C, was toxic on HeLa, Myeloma and T47D cell lines, with the LC50 of 4.3 μg/mL, 7.4 μg/mL and 27.8 μg/mL, respectively. The MJ-C was more potent than the basic protein MJ-30.

Keyword : acidic RIP, cytotoxic, Mirabilis jalapa,


Abstrak
Pada umumnya pemurnian Ribosome-inactivating protein (RIP) dengan menggunakan suatu kromatografi penukar kation sehingga diperoleh protein yang bersifat basa, seperti misalnya MJ-30 yang diisolasi dari daun Mirabilis jalapa. Fraksi protein M.jalapa yang tidak terikat pada kolom tersebut ternyata juga menunjukkan aktivitas pemotongan DNA superkoil dan bersifat sitotoksik terhadap beberapa sel kanker. Oleh karena itu protein tersebut perlu dimurnikan dan dikarakterisasi. Protein total diperoleh dengan mengendapkan ekstrak daun M.jalapa dengan amonium sulfat sampai jenuh. Setelah dilakukan dialisis, larutan protein tersebut dilewatkan kolom Ionenaustaucher Typ II, suatu penukar anion. Protein yang terikat dielusi dengan NaCl bergadien dari 0,0 – 0,5 M dalam bufer fosfat. Fraksi aktif ditentukan menggunakan uji aktivitas pemotongan DNA superkoil dan selanjutnya diuji aktivitas sitotoksisnya. Hasil penelitian menunjukkan bahwa fraksi yang mampu memotong DNA superkoil terelusi pada 0,35-0,40 M NaCl. Protein asam, MJ-C, bersifat sitotoksik terhadap sel HeLa, myeloma dan T47D dengan harga LC50 berturut-turut 14,3 μg/mL, 7,4 μg/mL dan 27,8 μg/mL. Ternyata MJ-C lebih poten dari pada MJ-30 yang besifat basa.

Kata kunci : RIP asam, sitotoksik, Mirabilis jalapa

Categories: Journal
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