Structural biology workflow for the expression and characterization of functional human sodium glucose transporter type 1 in Pichia pastoris.

ABSTRACT: Heterologous expression of human membrane proteins is a challenge in structural biology towards drug discovery. Here we report a complete expression and purification process of a functional human sodium/D-glucose co-transporter 1 (hSGLT1) in Pichia pastoris as representative example of a useful strategy for any human membrane protein. hSGLT1 gene was cloned in two different plasmids to develop parallel strategies: one which includes green fluorescent protein fusion for screening optimal conditions, and another for large scale protein production for structural biology and biophysics studies. Our strategy yields at least 1 mg of monodisperse purified recombinant hSGLT1 per liter of culture, which can be characterized by circular dichroism and infrared spectroscopy as an alpha-helical fold pr

SUBMITTER: Suades A 

PROVIDER: S-EPMC6362292 | biostudies-literature | 2019 Feb

REPOSITORIES: biostudies-literature