Sains Malaysiana 45(12)(2016): 1947–1957

http://dx.doi.org/10.17576/jsm-2016-4512-20

 

Pencirian Molekul Glikogen Sintase Kinase-3 dari Eimeria tenella

(Molecular Characterisation of Glycogen Synthase Kinase-3 from Eimeria tenella)

 

PING-PING YAO, MOHD FIRDAUS RAIH, HASIDAH MOHD SIDEK, NOOR EMBI

 & KIEW-LIAN WAN*

 

Pusat Pengajian Biosains & Bioteknologi, Fakulti Sains & Teknologi, Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor Darul Ehsan, Malaysia

 

Diserahkan: 14 April 2016/Diterima: 26 Oktober 2016

 

ABSTRAK

Penemuan sasaran dadah antikoksidia baharu merupakan antara usaha yang diperlukan untuk mengawal penyakit koksidiosis ayam yang disebabkan oleh spesies Eimeria. Dalam kajian ini, serpihan yang mengekodkan glikogen sintase kinase-3 (GSK-3) Eimeria tenella putatif telah diamplifikasi daripada cDNA E. tenella. Hasil pemadanan homologi menunjukkan jujukan GSK-3 E. tenella yang terjana mempunyai padanan yang tinggi dengan jujukan GSK-3 organisma lain. Domain terpulihara GSK-3 dan residu yang penting untuk aktiviti GSK-3 juga diramalkan hadir dalam jujukan GSK-3 E. tenella. Analisis struktur sekunder serta pemodelan homologi menunjukkan pembahagian struktur protein kepada domain bebenang beta pada hujung N dan domain heliks alfa pada hujung C, yang merupakan ciri enzim GSK-3. Kesemua hasil analisis ini menyokong bahawa jujukan yang dikaji mengekodkan protein GSK-3 dalam E. tenella. Walaupun darjah keterpuliharaan adalah tinggi, namun terdapat perbezaan yang bermakna diperhatikan antara GSK-3 E. tenella dan perumahnya. Residu Ser 9 yang dilaporkan penting untuk perencatan aktiviti GSK-3 didapati tidak terpulihara dalam GSK-3 E. tenella. Memandangkan Ser 9 merupakan tapak pemfosfatan bagi GSK-3β dalam haiwan vertebrata, ketiadaan residu ini dalam jujukan GSK-3 E. tenella mencadangkan bahawa pengawalaturan GSK-3 E. tenella melibatkan tapak pemfosfatan dan mekanisme yang berbeza. Tambahan pula, hasil analisis filogenetik menunjukkan bahawa GSK-3 E. tenella mempunyai pertalian yang rapat dengan protein GSK-3 tumbuh-tumbuhan. Analisis superposisi GSK-3 E. tenella dengan GSK-3β Homo sapiens pula menunjukkan bahawa perencat GSK-3 mampu berinteraksi dengan protein GSK-3 E. tenella. Keputusan kajian ini mencadangkan bahawa GSK-3 E. tenella mempunyai potensi untuk diperkembangkan sebagai sasaran dadah antikoksidia.

 

Kata kunci: Koksidiosis; parasit protozoa; sasaran dadah anti-koksidia

 

ABSTRACT

The discovery of new anticoccidial drug targets is amongst the necessary efforts needed to control chicken coccidiosis caused by Eimeria species. In this study, the fragment coding for the putative Eimeria tenella glycogen synthase kinase-3 (GSK-3) was amplified from the cDNA of E. tenella. Homology search showed that generated E. tenella GSK-3 sequence has high similarities with GSK-3 sequences from other organisms. The conserved domains of GSK-3 and residues important for the GSK-3 activity were also predicted within the E. tenella GSK-3. Secondary structure analysis and homology modelling predicted that the protein structure is divided into a beta strand domain at the N terminal and an alpha helix domain at terminal C, which are characteristics of GSK-3 enzymes. These results supported the E. tenella GSK-3 codes for the GSK-3 protein in E. tenella. Although the degree of conservation is high, significant differences were observed between GSK-3 of E. tenella and its host. The Ser 9 residue reported to be important for the inhibition of the GSK-3 activity was not conserved within the E. tenella GSK-3. Considering that Ser 9 is a phosphorylation site in GSK-3β of vertebrates, the absence of this residue in the E. tenella GSK-3 sequence suggests that the regulation of E. tenella GSK-3 involves a different phosphorylation site and mechanism. Phylogenetic analysis suggests that E. tenella GSK-3 has a closer relationship to plant GSK-3. Superposition analysis between E. tenella GSK-3 and Homo sapiens GSK-3β predicted that E. tenella GSK-3 is able to interact with a GSK-3 inhibitor. Taken together, these results suggested that the E. tenella GSK-3 has the potential to be developed into an anticoccidial drug target.

 

Keywords: Anti-coccidial drug target; coccidiosis; protozoan parasite

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*Pengarang untuk surat-menyurat; email: klwan@ukm.edu.my