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

Autor: Jed Jebali, José Luis, Assou el Battari, Amine Bazaa, Maram Morjen, Naziha Marrakchi, Sylvie Mathieu, Charlotte Jeanneau, Ali Gargouri, Mohamed El Ayeb
Přispěvatelé: Laboratoire des Venins et Toxines, Institut Pasteur de Tunis, Institut Pasteur de Tunis, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP), Laboratoire de Valorisation de la Biomasse et Production de Protéines chez les Eucaryotes [Sfax, Tunisie], Centre de Biotechnologie de Sfax (CBS), Centre de Recherches en Oncologie biologique et Oncopharmacologie (CRO2), Aix Marseille Université (AMU)- Hôpital de la Timone [CHU - APHM] (TIMONE)-Institut National de la Santé et de la Recherche Médicale (INSERM), Réseau International des Instituts Pasteur (RIIP), Faculté de Médecine de Tunis, Université de Tunis El Manar (UTM), The authors like to express their gratitude to Mr Anouar Smaoui from the English Language Unit at the Faculty of Science of Sfax, Tunisia, for his valuable help with the proofreading and language polishing of the present article.
Jazyk: angličtina
Rok vydání: 2012
Předmět:
Integrins
C‐type lectin
MESH: Viper Venoms/biosynthesis
MESH: Cricetinae
Kidney
MESH: Lectins
C-Type/chemistry
MESH: Kidney/cytology
law.invention
MESH: Cell Movement/drug effects
MESH: Cell Proliferation/drug effects
MESH: Cricetulus
Cell Movement
C-type lectin
law
Cricetinae
lebecetin
MESH: Recombinant Proteins/biosynthesis
MESH: Animals
snake venom
MESH: Viper Venoms/genetics
0303 health sciences
biology
030302 biochemistry & molecular biology
Transfection
MESH: Protein Subunits
MESH: Kidney/metabolism
MESH: Viper Venoms/chemistry
Recombinant Proteins
MESH: Integrins/metabolism
MESH: Recombinant Proteins/chemistry
MESH: HEK293 Cells
Recombinant DNA
Biotechnology
MESH: Cell Line
Tumor
integrin
Protein subunit
Blotting
Western
CHO Cells
Viper Venoms
03 medical and health sciences
Cricetulus
Affinity chromatography
MESH: CHO Cells
MESH: Viper Venoms/pharmacology
Cell Line
Tumor
Complementary DNA
Animals
Humans
MESH: Blotting
Western
Lectins
C-Type
MESH: Recombinant Proteins/genetics
MESH: Recombinant Proteins/pharmacology
Cell Proliferation
030304 developmental biology
MESH: Humans
heterodimerization
MESH: Lectins
C-Type/genetics
Lectin
Embryo
Mammalian
Molecular biology
Protein Subunits
HEK293 Cells
Glycoprotein Ib
biology.protein
MESH: Lectins
C-Type/biosynthesis
MESH: Embryo
Mammalian/cytology
[SDV.MHEP]Life Sciences [q-bio]/Human health and pathology
Zdroj: Biotechnology Progress
Biotechnology Progress, Wiley, 2012, 28 (6), pp.1560-1565. ⟨10.1002/btpr.1632⟩
ISSN: 8756-7938
DOI: 10.1002/btpr.1632⟩
Popis: International audience; 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.
Databáze: OpenAIRE
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