Abstract
Triosephosphate isomerase (TIM) is a major enzyme in the glycolytic pathway, which catalyzes the interconversion of glyceraldehyde 3-phosphate to dihydroxyacetone phosphate. Here, we report cloning, expression and purification of a catalytically active recombinant TIM of Leishmania donovani (LdTIM). The recombinant LdTIM had a pH optimum in the range of 7.2-9.0, found stable at 25°C for 30 min and K(m) and V(max) for the substrate glyceraldehyde 3-phosphate was 0.328±0.02mM and 10.05mM/min/mg, respectively. The cysteine-reactive agent methylmethane thiosulphonate (MMTS) was used as probe, in order to test its effect on enzyme activity. The MMTS induced 75% enzyme inactivation within 15 min at 250 μM concentration. The biochemical characterization of LdTIM described in this work is the essential step towards deeper understanding of its role in parasite survival. The purification of LdTIM in bioactive form provides important tools for further functional and structural studies.
References
Jan 1, 1988·Biology of the Cell·P A Michels
Mar 1, 1994·European Journal of Biochemistry·L KohlP A Michels
Feb 1, 1993·European Journal of Biochemistry·T V BorchertR K Wierenga
Dec 1, 1995·Chemistry & Biology·A Gómez-PuyouR Pérez-Montfort
Oct 1, 1996·European Journal of Biochemistry·G Garza-RamosA Gómez-Puyou
Mar 15, 1997·European Journal of Biochemistry·P Ostoa-SalomaR Pérez-Montfort
Jul 1, 1997·European Journal of Biochemistry·A LandaD A Fernández-Velasco
Jul 8, 1998·European Journal of Biochemistry·G Garza-RamosA Gómez-Puyou
May 11, 1999·Protein Engineering·J C WilliamsR K Wierenga
Jul 9, 1999·The Journal of Infectious Diseases·R LiraD Sacks
Sep 12, 2000·International Journal for Parasitology·L JiménezA Landa
Oct 26, 2000·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·S SundarH W Murray
Feb 17, 2001·International Journal of Infectious Diseases : IJID : Official Publication of the International Society for Infectious Diseases·H W Murray
May 15, 2002·The Journal of Biological Chemistry·Kapil MaithalPadmanabhan Balaram
Nov 19, 2003·European Journal of Biochemistry·Héctor VillaDavid Ordóñez
Nov 13, 2004·FEMS Microbiology Reviews·Juliette MoyersoenPaul A M Michels
Sep 19, 2006·FEMS Microbiology Letters·Divya MathurLalit C Garg
Dec 14, 2007·Molecular and Biochemical Parasitology·Sergio Enriquez-FloresHoracio Reyes-Vivas
Apr 11, 2008·Clinical Microbiology Reviews·Jorge AlvarJavier Moreno
Aug 28, 2009·Expert Opinion on Emerging Drugs·Margriet Leontine den BoerManica Balasegaram
Aug 21, 2010·FEMS Microbiology Letters·Prachi BhargavaUma Roy
Mar 3, 2011·Applied Biochemistry and Biotechnology·Kishore KumarUma Roy
Citations
Aug 27, 2013·Biochimie·Veronika L ZinsserDavid J Timson
Jul 5, 2013·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Ifedayo Victor Ogungbe, William N Setzer
Oct 21, 2016·Biochimica Et Biophysica Acta·Alonso A Lopez-ZavalaLuis G Brieba
Jan 28, 2017·Developmental and Comparative Immunology·Fei LiuFuhua Li
Dec 28, 2019·Microorganisms·Beatriz Hernández-OchoaEdgar Sierra-Palacios
Sep 29, 2020·Frontiers in Immunology·Xuzheng LiaoShuang Zhang
Nov 5, 2014·Chemical Reviews·Advait S NagleValentina Molteni