Nucleoside diphosphate kinases (NDK; EC 184.108.40.206) are enzymes required for maintaining
intracellular levels of nucleosides triphosphates (NTP) through transfer the -phosphoryl group from a
NTP to a NDP. The enzyme is associated with several biological functions including prevention of
host ATP-mediated cytolysis during pathogenic infections. Here we present the biophysical characterization
of NDK from Leishmania major and the effect of a mutation on the protein structure in solution.
The structural stability was analyzed since this secreted protein may act in different microenvironments at various stages
of the parasite life cycle. LmNDK and P95S mutant were subjected to denaturation with pH and guanidine. Structural
transitions were monitored by circular dichroism and intrinsic fluorescence tryptophan emission. Our results showed that
the LmNDK is more structurally stable than other described NDKs and that the catalytically active P95S mutant in the
Kpn loop presented a decrease in protein stability, indicating the importance of this proline for maintenance of the
Keywords: Drug-target protein, leishmaniasis, nucleoside diphosphate kinase, nucleotide metabolism, protein biophysical
characterization, protein stability.
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