Cloning, purification, and homology modeling of Histone deacetylase in Leishmania donovani

• Published in: Infection, genetics and evolution genetics and evolution, 2021-04, Vol.89, p.104738-104738, Article 104738
• Main Authors: Prasanna, Pragya, Kumar, Rakesh, Singh, Vijay Kumar, Upadhyay, Arun
• Format: Article
• Language: English
• Subjects: Chromatography, Liquid; Cloning, Molecular; Drug target; Histone deacetylase; Histone Deacetylases - genetics; Histone Deacetylases - isolation & purification; Histone Deacetylases - metabolism; Homology modeling; Kala-azar; Leishmania donovani; Leishmania donovani - enzymology; Molecular Dynamics Simulation
• ISSN: 1567-1348; 1567-7257
• DOI: 10.1016/j.meegid.2021.104738

Neglected diseases, such as leishmaniasis, are still a major health problem in poor countries. To date, there is a severe lack of effective, safe, and affordable treatment for leishmaniasis. Currently, there are very limited chemotherapeutic options, and the development of vaccines is still underway. Hence, novel therapeutic strategies need to be developed against leishmanial parasites. Histone deacetylases (HDACs), silent regulators of many critical pathways, have been validated as potential therapeutic targets in cancer and several parasitic diseases. In the present work, we have isolated and characterized biologically active Zn2+-dependent HDAC protein from leishmania that can be studied further as a potential anti-leishmanial drug target to develop new therapies against neglected diseases. The nucleotide sequence of the HDAC gene with no intervening sequence was amplified, cloned in a pET-28a vector, and later transformed into the BL21(DE3) competent E. coli bacterial cells. After transformation, the cells were cultured and induced with 0.6 mM of IPTG to express histidine-tagged HDAC protein (LD_HDAC), which was later purified using nickel affinity chromatography. The approximate protein size confirmed with the help of 10% SDS–PAGE was ~48.0 kDa. The enzymatic assay using the purified protein confirmed it as biologically active. A three dimensional structure of LD_HDAC was modeled using the crystal structure of HDAC2 protein of Homo sapiens (PDB ID: 6G3O). This protein can be utilized for the screening of Leishmania-specific HDAC inhibitors. [Display omitted] •Leishmaniasis, a fatal disease remains a major public health concern in developing countries.•Current therapeutic strategies are insufficient to prevent the infection and pathogenesis.•Histone deacetylases, a critical regulator of several pathways is essential for parasite survival.•Characterization of HDAC gene in Leishmania donovani has not been achieved yet.•Characterization of parasite HDAC proteins may provide novel insights for developing therapeutic strategies.