Expression, purification and spectrophotometric analysis of nucleoside hydrolase from Leishmania chagasi (LcNH).

Abstract

Leishmaniasis represents an important public health problem in several countries. The main target in this study is the nucleoside hydrolase Leishmania chagasi (LcNH) that is responsible for causing visceral leishmaniasis, principally in Brazil. Nucleoside hydrolase enzymes are members of this pathway, hydrolyzing the N-glycosidic bond of ribonucleosides for the synthesis of nucleic acids. We present here for the first time, the expression and purification protocols to obtain the enzymes LcNH1 and LcNH2 that can be employed to explore novel strategies to produce nucleoside hydrolase inhibitors for use in chemotherapy. Protein integrity was also confirmed by SDS-PAGE gel, mass spectrometry and enzymatic activity.

• Authors: Edijane M Sales, Gileno S Sousa, Chiraz Belouezzane, Fábio C L Almeida, José D Figueroa-Villar
• Journal: Protein expression and purification (Protein Expr Purif) Vol. 161 Pg. 40-48 (09 2019) ISSN: 1096-0279 [Electronic] United States
• PMID: 31034876 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2019 Elsevier Inc. All rights reserved.

Chemical References

• Protozoan Proteins
• N-Glycosyl Hydrolases

Topics

• Cloning, Molecular
• Leishmania (enzymology, genetics)
• Magnetic Resonance Spectroscopy
• Mass Spectrometry
• N-Glycosyl Hydrolases (chemistry, genetics, isolation & purification, metabolism)
• Protozoan Proteins (chemistry, genetics, isolation & purification, metabolism)