TITLE:
Identification and Characterization of Heparan Sulphate Binding Proteins of Entamoeba histolytica
AUTHORS:
Upninder Kaur, Sumeeta Khurana, Uma Nahar Saikia, Mohan Lal Dubey, Rakesh Sehgal
KEYWORDS:
Heparan Sulphate, Entamoeba histolytica, Identification, Characterization
JOURNAL NAME:
Journal of Biosciences and Medicines,
Vol.6 No.8,
August
9,
2018
ABSTRACT: A large
number of microbial pathogens bind to heparan sulphate on eukaryotic cell
surfaces. Heparan Sulphate Binding Proteins (HSBPs) from Entamoeba histolytica culture lysates were obtained by sequential
ammonium sulphate precipitation and Protein purify. SDS-PAGE and immunoblotting
experiments indicated the presence of two major extracellular proteins in E. histolytica (51.2 kDa and 61.0 kDa). Characterization
of HSBPs by 2D Gel electrophoresis of 40% (NH4)2SO4 precipitated lysate of E. histolytica revealed that the isoelectric point of 51.2 kDa HSBP was at pH3.0. The
protein of 61.0 kDa HSBP showed three spots in 40% (NH4)2SO4 lysate which had isoelectric point between pH 4.0 - 7.0. While in 80% (NH4)2SO4 precipitated lysate, 51.2 kDa HSBP showed only one spot which had isoelectric
point at pH 3. However, 61.0 kDa
HSBP revealed 2 spots which had isoelectric point between pH 4 and 5. The
result showed that this parasite has proteins which interact with heparan
sulphate whose molecular formula is C14H23NO21S-23. These proteins may have a role in binding of
parasite to heparan sulphate on host cells. Further characterization by MALDI-TOF
analysis of HSBPs from E. histolytica demonstrated HSBPs to be novel protein in this parasite which has been
isolated, purified and characterized first time by our group in the present
study.