Establishment of correlation between in-silico and in-vitro test analysis against Leishmania HGPRT to inhibitors

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• 作者：Md Yousuf Ansari^a ; ^b ; Asif Equbal^c ; Manas Ranjan Dikhit^b ; Rani Mansuri^a ; ^b ; Sindhuprava Rana^b ; Vahab Ali^c ; Ganesh Chandra Sahoo^b ; ^{ganeshiitkgp@gmail.com" class="auth_mail" title="E-mail the corresponding author} ; ^{ganeshcs@icmr.org.in" class="auth_mail" title="E-mail the corresponding author} ; Pradeep Das^a ; ^b
• 关键词：HGPRT ; hypoxanthine-guanine phosphoribosyltransferase ; HPRT ; hypoxanthine phosphoribosyltransferase ; APRT ; adenosine phosphoribosyltransferase ; XPRT ; xanthine phosphoribosyltransferase ; CL ; cutaneous leishmaniasis ; MCL ; muco-cutaneous leishmaniasis ; VL ; visceral leishmaniasis ; PKDL ; post-Kala-azar dermal leishmaniasis ; BLAST ; basic local alignment search tool ; PDB ; protein data bank ; MTT ; 3-(4 ; 5-dimethylthiazol-2-yl)-2 ; 5-diphenyltetrazolium bromide ; RMSD ; root mean square deviation ; GMP ; guanosi
• 刊名：International Journal of Biological Macromolecules
• 出版年：2016
• 出版时间：February 2016
• 年：2016
• 卷：83
• 期：Complete
• 页码：78-96
• 全文大小：5812 K

文摘

Hypoxanthine Phosphoribosyltransferase (HGPRT; EC 2.4.2.8) is a central enzyme in the purine recycling pathway of all protozoan parasites. Protozoan parasites cannot synthesize purine bases (DNA/RNA) which is essential for survival as lack of de-novo pathway. Thus its good target for drug design and discovery as inhibition leads to cessation of replication. PRTase (transferase enzyme) has common PRTase type I folding pattern domain for its activities. Genomic studies revealed the sequence pattern and identified highly conserved residues that catalyzed the reaction in protozoan parasites. A recombinant protein has 24 kDa molecular mass (rLdHGPRT) was cloned, expressed and purified for testing of guanosine monophosphate (GMP) analogous compounds in-vitro by spectroscopically to the rLdHGPRT, lysates protein and MTT assay on Leishmania donovani. The predicted inhibitors of different libraries were screen into FlexX. The reported inhibitors were tested in-vitro. The 2′-deoxyguanosine 5′-diphosphate (DGD) (IC₅₀ value 12.5 μM) is two times more effective when compared to guanosine-5′-diphosphate sodium (GD). Interestingly, LdHGPRT complex has shown stable after 24 ns in molecular dynamics simulation with interacting amino acids are Glu125, Ile127, Lys87 and Val186. QSAR studies revealed the correlation between predicted and experimental values has shown R² 0.998. Concludes that inversely proportional to their docked score with activities.