title: Purification and characterization of guanylate kinase, a nucleoside monophosphate kinase of Brugia malayi.
abstract: Guanylate kinase, a nucleoside monophosphate kinase of Brugia malayi which is involved in reversible transfer of phosphate groups from ATP to GMP, was cloned, expressed and characterized. The native molecular mass of BmGK was found to be 45 kDa as determined by size exclusion chromatography and glutaraldehyde cross-linking which revealed that the protein is homodimer in nature. This is a unique characteristic among known eukaryotic GKs. GMP and ATP served as the most effective phosphate acceptor and donor, respectively. Recombinant BmGK utilized both GMP and dGMP, as substrates showing Km values of 30 and 38 μ m, respectively. Free Mg+2 (un-complexed to ATP) and GTP play a regulatory role in catalysis of BmGK. The enzyme showed higher catalytic efficiency as compared with human GK and showed ternary complex (BmGK-GMP-ATP) formation with sequential substrate binding. The secondary structure of BmGK consisted of 45% α-helices, 18% β-sheets as revealed by CD analysis. Homology modelling and docking with GMP revealed conserved substrate binding residues with slight differences. Differences in kinetic properties and oligomerization of BmGK compared with human GK can provide the way for design of parasite-specific inhibitors.
authors: Division of Biochemistry,CSIR-Central Drug Research Institute,Lucknow - 226031, Uttar Pradesh,India.
authors: Smita
authors: S
authors: Gupta
authors: Division of Biochemistry,CSIR-Central Drug Research Institute,Lucknow - 226031, Uttar Pradesh,India.
authors: Sunita
authors: S
authors: Yadav
authors: Division of Molecular and Structural Biology,CSIR-Central Drug Research Institute,Lucknow - 226031, Uttar Pradesh,India.
authors: Nidhi
authors: N
authors: Singh
authors: Division of Biochemistry,CSIR-Central Drug Research Institute,Lucknow - 226031, Uttar Pradesh,India.
authors: Anita
authors: A
authors: Verma
authors: Division of Biochemistry,CSIR-Central Drug Research Institute,Lucknow - 226031, Uttar Pradesh,India.
authors: Imran
authors: I
authors: Siddiqi
authors: Division of Biochemistry,CSIR-Central Drug Research Institute,Lucknow - 226031, Uttar Pradesh,India.
authors: Jitendra K
authors: JK
authors: Saxena
mesh_terms: D000818:Animals; D017178:Brugia malayi / Q000235:genetics; D019281:Dimerization; D005368:Filariasis / Q000469:parasitology*; D051528:Guanylate Kinases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008958:Models, Molecular; D008970:Molecular Weight; D009703:Nucleoside-Phosphate Kinase / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D009703:Nucleoside-Phosphate Kinase; D051528:Guanylate Kinases