Minor Impact of A258D Mutation on Biochemical and Enzymatic Properties of Leishmania infantum GDP-Mannose Pyrophosphorylase

Abstract

Background: Leishmaniasis, a vector-borne disease caused by the protozoan parasite from the genus Leishmania, is endemic to tropical and subtropical areas. Few treatments are available against leishmaniasis, with all presenting issues of toxicity, resistance, and/or cost. In this context, the development of new antileishmanial drugs is urgently needed. GDP-mannose pyrophosphorylase (GDP-MP), an enzyme involved in the mannosylation pathway, has been described to constitute an attractive therapeutic target for the development of specific antileishmanial agents. Methods: In this work, we produced, purified, and analyzed the enzymatic properties of the recombinant L. infantum GDP-MP (LiGDP-MP), a single leishmanial GDP-MP that presents mutation of an aspartate instead of an alanine at position 258, which is also the single residue difference with the homolog in L. donovani: LdGDP-MP. Results: The purified LiGDP-MP displayed high substrate and cofactor specificities, a sequential random mechanism of reaction, and the following kinetic constants: Vm at 0.6 µM·min−1, Km from 15–18 µM, kcat from 12.5–13 min−1, and kcat/Km at around 0.8 min−1µM−1. Conclusions: These results show that LiGDP-MP has similar biochemical and enzymatic properties to LdGDP-MP. Further studies are needed to determine the advantage for L. infantum of the A258D residue change in GDP-MP.

Antileishmanial and Antitrypanosomes Drugs for the Current Century

“…No substantial modifications were found when compared to the homolog molecule from anthroponotic L. donovani (LdGDP-MP). Therefore, the potential advantage of modifying residue 258 for L. infantum should be investigated [ 39 ]. The mechanistic explanation of the resistance phenomena is strongly linked to the MoA, but our knowledge is largely incomplete.…”

The Potential of 2-Substituted Quinolines as Antileishmanial Drug Candidates

“…GDP-Mannose Pyrophosphorylase (GDP-MP) is an enzyme involved in host–parasite recognition considered to be essential for parasite infection [ 83 , 84 ]. GDP-MPs were purified from L. mexicana ( Lm GDP-MP) and L. donovani ( Ld GDP-MP), and their enzymatic properties were compared with the human enzyme ( h GDP-MP) [ 60 , 85 , 86 , 87 ]. From a rationale design strategy including molecular modeling of 100 potential inhibitors, four compounds were identified as having a promising and specific inhibitory effect on parasite GDP-MP associated with antileishmanial activity.…”