Expression of a functional recombinant C‐type lectin‐like protein lebecetin in the human embryonic kidney cells |
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| Authors: | Jed Jebali Sylvie Mathieu Amine Bazaa Mohamed el Ayeb José Luis Ali Gargouri Naziha Marrakchi Assou el Battari |
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| Institution: | 1. Laboratoire des Venins et Toxines, Institut Pasteur de Tunis, Tunisie;2. Laboratoire de Valorisation de la Biomasse et Production de Protéines chez les Eucaryotes, Centre de la Biotechnologie de Sfax (CBS);3. Centre de Recherche en Oncologie biologique et Oncopharmacologie (CRO2), INSERM UMR 911, Marseille, France;4. Aix‐Marseille Université, Marseille, France;5. Faculté de Médecine de Tunis, Tunisie |
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| 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. © 2012 American Institute of Chemical Engineers Biotechnol. Prog., 2012 |
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| Keywords: | snake venom lebecetin C‐type lectin heterodimerization integrin |
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