Generation of nanobodies against different targets (three membrane proteins) for structural studies

FERNANDO A GOLDBAUM; ALZOGARAY V

Bruselas

Workshop; Nanobodies for Structural Biology and beyond; 2018

VIB-VUB center for Structural Biology, Vrije Universiteit Brussel.

Currently, I am working in the setup of a local platform to generate Nb against a few membrane proteins of relevance in human medicine with unknown 3D structure, as follows:(i) Voltage activated sodium channels (Nav), which consist of a major subunit (α) whose activity is modulated by several channel interacting proteins (CIP). These channels are the major participants in the depolarization and electrical conductance of neurons, muscle cells and heart cells (Gabelli SB et al 2016), (ii) Phosphoinositide 3-kinases (PI3Ks), which are ubiquitous lipid kinases that activate signaling cascades controlling cell survival, proliferation, protein synthesis, and vesicle trafficking (Maheshwari S et al 2017), (iii) Na+/I− symporter (NIS), which is a key plasma membrane glycoprotein in thyroid gland. NIS transports different substrates with different stoichiometries. Furthermore, it is increasingly used as a highly effective reporter gene for imaging techniques (Ravera S et al 2017), and (iv) The histidine kinase (HK), DesK, from Bacillus subtilis, which is a membrane-bound thermosensor suited to remodel membrane fluidity when the ambient temperature drops below approximately 30 °C. These cold sensors are ubiquitous integral membrane proteins found in all kingdoms of life. They are involved in many physiological roles, including membrane remodeling, chemotaxis, touch, and pain (Saita E et al 2015).