Production and functional characterization of collectin-11 mutants associated with 3MC syndrome

VIDELA GILETTA, MARÍA BELÉN; BECCACECE, IGNACIO; CARRIZO, MARÍA E.

Mar del Plata, Buenos Aires

Congreso; LI Reunión Anual de la Sociedad Argentina de Investigación Bioquímica y Biología Molecular (SAIB); 2015

Sociedad Argentina de Investigación Bioquímica y Biología Molecular (SAIB)

C-type lectins are proteins that bind carbohydrates in a Ca2+-dependent manner. Collectins are C-type lectins that contain a collagen-like domain. Their basic functional unit is a trimer whose monomeric subunits have, besides the collagen-like region, a coiled-coil neck domain and a C-type lectin domain, also referred to as carbohydrate recognition domain (CRD). Collectin-11 (CL-11) was initially identified in a library of human kidney, so it is also known as collectin kidney or CL-K1. It binds fucose and more weakly mannose, as well as polysaccharides isolated from certain microorganisms. CL-11 would also bind plasmid DNA and bacterial and mammalian genomic DNA. Besides, it has been defined as a new molecule in the lectin pathway of the complement associated MASPs system. Mutations in the genes that encode MASP-3 and CL-11 cause a rare genetic disorder named 3MC syndrome, that exhibits a characteristic facial dysmorphism, growth deficiency, learning disability, genital, limb and renal anomalies and, in some cases, heart abnormalities and other skeletal malformations. Here we describe the method we used to obtain the fragment comprising the neck and the CRD of wild type CL-11 and of three mutants associated to 3MC syndrome, after refolding from inclusion bodies, and present an analysis of their ability to bind ligands.