title: Molecular and functional characterisation of a stress responsive cysteine protease, EhCP6 from Entamoeba histolytica.
abstract: Entamoeba histolytica cysteine protease 6 (EhCP6) is a stress responsive cysteine protease that is upregulated in response to heat shock and during pathogen invasion of the host tissue. In the present study an attempt has been made to express and purify recombinant EhCP6 in order to gain insights into its biochemical properties. The recombinant and refolded protein has been shown to undergo autoproteolysis in the presence of DTT and SDS to give rise to ∼25kDa mature form. The mature form of the protein was found to exhibit a protease activity that is sensitive to E-64, a specific cysteine protease inhibitor. In silico homology modelling of EhCP6 revealed that the protein exhibits conservation of almost all the major structural features of cathepsin-L like cysteine proteases. Further in vivo studies are needed to decipher the function of the protein in response to different stressed conditions.
authors: Division of Plant Biology, Bose Institute, Centenary Campus, P1/12 C.I.T Road, Scheme VIIM, Kolkata 700054, India. Electronic address: ghosh.anupama@jcbose.ac.in.
authors: Anupama
authors: A
authors: Ghosh
authors: Integrated Science Education and Research Center (ISERC) and Department of Biotechnology, Visva-Bharati University, Santiniketan 731235, India.
authors: Sanghamitra
authors: S
authors: Raha
mesh_terms: D000595:Amino Acid Sequence; D057056:Cysteine Proteases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D004748:Entamoeba histolytica / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D015800:Protozoan Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D013312:Stress, Physiological; D040681:Structural Homology, Protein; D013696:Temperature
chemical_list: D015800:Protozoan Proteins; D057056:Cysteine Proteases
keywords: Cysteine protease; Entamoeba histolytica; Stress response