Expression of a functional recombinant C-type lectin-like protein lebecetin in the human embryonic kidney (HEK) cells. - Archive ouverte HAL Access content directly
Journal Articles Biotechnology Progress Year : 2012

Expression of a functional recombinant C-type lectin-like protein lebecetin in the human embryonic kidney (HEK) cells.

(1, 2) , (3) , (4) , (3) , (1) , (1) , (3) , (2) , (1, 5) , (3)
1
2
3
4
5

Abstract

Lebecetin is an anticoagulant C-type lectin-like protein (CLPs) that was previously isolated from Macrovipera lebetinavenom and described to consist of two subunits (alpha and beta). It was reported to potently prevent platelet aggregation by binding to glycoprotein Ib (GPIb) and to exhibit a broad spectrum of inhibitory activities on various integrin-mediated functions of tumour cells, including adhesion, proliferation, and cell migration. The present study aimed to investigate the structure-function of lebecetin. Accordingly, the cDNA of each subunit was cloned and separately or jointly expressed in the human embryonic kidney cells (HEK)using two vectors with different selectable tags. The immunofluorescence analysis of transfected cells revealed significant expression levels and co-localization of the two lebecetin subunits. The recombinant proteins were efficiently secreted and purified using metal-chelating affinity chromatography. We found that the Lebecetin alpha and beta subunits were produced as a mixture of homodimers and heterodimers and that the heterodimerization represent a prerequisite for functioning.

Dates and versions

pasteur-00733000 , version 1 (17-09-2012)

Identifiers

Cite

Jed Jebali, Charlotte Jeanneau, Maram Morjen, Sylvie Mathieu, Amine Bazaa, et al.. Expression of a functional recombinant C-type lectin-like protein lebecetin in the human embryonic kidney (HEK) cells.. Biotechnology Progress, 2012, 28 (6), pp.1560-1565. ⟨10.1002/btpr.1632⟩. ⟨pasteur-00733000⟩
38 View
0 Download

Altmetric

Share

Gmail Facebook Twitter LinkedIn More