Heterologous expression and biochemical characterization of the recombinant nucleoside triphosphate diphosphohydrolase 2 (LbNTPDase2) from Leishmania (Viannia) braziliensis

被引：0
作者：
Pavione, Nancy da Rocha Torres [1 ,2 ]
de Moraes, Joao Victor Badaro [1 ,2 ]
Ribeiro, Isadora Cunha [1 ]
de Castro, Raissa Barbosa [1 ]
da Silva, Walmir [1 ]
de Souza, Anna Claudia Alves [1 ]
da Silva, Victor Hugo Ferraz [1 ]
Vasconcellos, Raphael de Souza [1 ]
Bressan, Gustavo da Costa [1 ]
Fietto, Juliana Lopes Rangel [1 ]
机构：
[1] Univ Fed Vicosa, Biochem & Mol Biol Dept, Vicosa, MG, Brazil
[2] Univ Fed Vicosa, Gen Biol Dept, Vicosa, MG, Brazil
来源：
PURINERGIC SIGNALLING | 2024年 / 20卷 / 05期
关键词：
Leishmania braziliensis; LbNTPDase2; Recombinant protein expression; Enzyme characterization; DEPENDENT ECTO-ATPASE; TOXOPLASMA-GONDII; BACTERIAL EXPRESSION; STRUCTURAL INSIGHT; TRYPANOSOMA-CRUZI; NTPDASE; AMAZONENSIS; INHIBITION; LOCALIZATION; PURIFICATION;
D O I：
10.1007/s11302-023-09980-9
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
Leishmania braziliensis is a pathogenic protozoan parasite that causes American Tegumentary Leishmaniasis (ATL), an important tropical neglected disease. ENTPDases are nucleotidases that hydrolyze intracellular and/or extracellular nucleotides. ENTPDases are known as regulators of purinergic signalling induced by extracellular nucleotides. Leishmania species have two isoforms of ENTPDase, and, particularly, ENTPDase2 seems to be involved in infectivity and virulence. In this study, we conducted the heterologous expression and biochemical characterization of the recombinant ENTPDase2 of L. braziliensis (rLbNTPDase2). Our results show that this enzyme is a canonical ENTPDase with apyrase activity, capable of hydrolysing triphosphate and diphosphate nucleotides, and it is dependent on divalent cations (calcium or magnesium). Substrate specificity was characterized as UDP>GDP>ADP>GTP>ATP=UTP. The enzyme showed optimal activity at a neutral to basic pH and was partially inhibited by suramin and DIDS. Furthermore, the low apparent Km for ADP suggests that the enzyme may play a role in adenosine-mediated signalling. The biochemical characterization of this enzyme can open new avenues for using LbNTPDase2 as a drug target.
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页码：509 / 520
页数：12
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