Abstract

This is auto test ....

The characterization of native protein assemblies requires the production of a relatively pure sample that maintains the full complement, native organization, and function of that complex. This can be particularly challenging to achieve for large, multi-component, membrane embedded complexes using the traditional recombinant expression and reconstitution methodologies. However, using affinity capture from native cells, suitable whole endogenous protein complexes can be isolated. In this webinar, I will present a method for the affinity isolation and characterization of baker's yeast (S. cerevisiae) nuclear pore complexes, which are ~50 MDa assemblies made up of 552 distinct proteins and embedded in a double-membraned nuclear envelope. Producing this sample allowed us to perform analyses to characterize the mass, stoichiometry, morphology, and connectivity of this complex and to obtain its integrative structure with subnanometer precision. We believe this methodology can be applied to other challenging protein complexes to produce similar results.

The speakers will discuss:

a) Affinity purification of large, native macromolecular assemblies

b) Integrative structural characterization of the nuclear pore complex