title: Expression of a functional recombinant C-type lectin-like protein lebecetin in the human embryonic kidney cells.
abstract: Lebecetin is an anticoagulant C-type lectin-like protein that was previously isolated from Macrovipera lebetina venom and described to consist of two subunits (alpha and beta). It was reported to potently prevent platelet aggregation by binding to glycoprotein Ib and to exhibit a broad spectrum of inhibitory activities on various integrin-mediated functions of tumor cells, including adhesion, proliferation, and cell migration. This 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 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 represents a prerequisite for functioning.
authors.affiliation: Laboratoire des Venins et Toxines, Institut Pasteur de Tunis, Tunisia.
authors.forename: Jed
authors.initials: J
authors.lastname: Jebali
authors.forename: Charlotte
authors.initials: C
authors.lastname: Jeanneau
authors.forename: Maram
authors.initials: M
authors.lastname: Morjen
authors.forename: Sylvie
authors.initials: S
authors.lastname: Mathieu
authors.forename: Amine
authors.initials: A
authors.lastname: Bazaa
authors.forename: Mohamed
authors.initials: M
authors.lastname: El Ayeb
authors.forename: José
authors.initials: J
authors.lastname: Luis
authors.forename: Ali
authors.initials: A
authors.lastname: Gargouri
authors.forename: Naziha
authors.initials: N
authors.lastname: Marrakchi
authors.forename: Assou
authors.initials: A
authors.lastname: el Battari
mesh_terms: D000818:Animals; D015153:Blotting, Western; D016466:CHO Cells; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D006224:Cricetinae; D003412:Cricetulus; D004622:Embryo, Mammalian / Q000166:cytology; D057809:HEK293 Cells; D006801:Humans; D016023:Integrins / Q000378:metabolism; D007668:Kidney / Q000166:cytology / Q000378:metabolism*; D037181:Lectins, C-Type / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D021122:Protein Subunits; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology; D014757:Viper Venoms / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology
chemical_list: D016023:Integrins; D037181:Lectins, C-Type; D021122:Protein Subunits; D011994:Recombinant Proteins; D014757:Viper Venoms; C478765:lebecetin protein, Macrovipera lebetina