title: Heterologous expression, purification and characterization of L-type lectin homologue fromLeishmania donovani.
abstract: Leishmaniasis, a disease of the developing world affects about 12 million people and has limited therapeutic interventions available. L-type lectins, Endoplasmic Reticulum Golgi Intermediate Compartment/Vesicular Integral Proteins (ERGIC-53/VIP36) are involved in protein sorting in luminal compartments of animal cells and are important for parasite biology. A lectin homologue was identified through a bioinformatics analysis ofLeishmaniagenome and it was found to have N-terminal conserved carbohydrate recognition domain (CRD) and a unique C-terminal region rich in repetitive amino acids and a poly glutamine tract. The N-terminal CRD region was cloned and expressed inEscherichia coli, but gave an insoluble expression which was re-solubilized by on column refolding. The fold integrity was checked through CD, fluorescence and functional assay of hemagglutination activity using rabbit erythrocyte. Bioinformatics analysis identified 15 members from Tritryps (Leishmaniaspp.,Trypanosomaspp.) and they separate out as a distinct clade in the global phylogenetic analysis of all ERGIC-53/VIP36 sequences downloaded from Uniprot. Our analysis shows that the extended C-terminal regions with repeats is unique to Tritryps and this repeat pattern is different in sequences fromLeishmaniaspp.andTrypanosomaspp.and all these features make this protein an interesting candidate for further detailed studies.
authors.affiliation: Bioinformatics & Structural Biology, Indian Institute of Advanced Research, Koba, Gandhinagar, 382007 Gujarat, India.
authors.forename: Vijay
authors.initials: V
authors.lastname: Singh
authors.affiliation: Bioinformatics & Structural Biology, Indian Institute of Advanced Research, Koba, Gandhinagar, 382007 Gujarat, India.
authors.forename: Divya N
authors.initials: DN
authors.lastname: Nair
authors.affiliation: Bioinformatics & Structural Biology, Indian Institute of Advanced Research, Koba, Gandhinagar, 382007 Gujarat, India.
authors.forename: Radhey Shyam
authors.initials: RS
authors.lastname: Kaushal
authors.affiliation: Bioinformatics & Structural Biology, Indian Institute of Advanced Research, Koba, Gandhinagar, 382007 Gujarat, India.
authors.forename: Manoj
authors.initials: M
authors.lastname: Kumar
authors.affiliation: Bioinformatics & Structural Biology, Indian Institute of Advanced Research, Koba, Gandhinagar, 382007 Gujarat, India.
authors.forename: Anju
authors.initials: A
authors.lastname: Pappachan
authors.affiliation: Bioinformatics & Structural Biology, Indian Institute of Advanced Research, Koba, Gandhinagar, 382007 Gujarat, India; Centre for Genetic Disease and Molecular Medicine, Central University of Punjab, Bathinda 151001, India.
authors.forename: Desh Deepak
authors.initials: DD
authors.lastname: Singh
keywords: Antigen; Hemagglutination; L-type lectin; Modeling