The Fall 2022 GWC Seminar Series will kick off Wednesday, September 21st. These seminars will be held in person in the Main Link room at the home University of the presenter, and via Teams for the opposite campus.
Exploring conformational dynamics of biological supramolecular machines using methyl-TROSY NMR spectroscopy
Assistant
Professor Rui
Huang
Department
of
Chemistry
University
of
Guelph
Wednesday,
September
21,
2022
3:00
p.m.
Main
link
room
at
the
University
of
Guelph
-
Coffee
and
snacks
served.
Online
via
Microsoft
Teams
for
the
University
of
Waterloo.
Please
contact
gwc@uoguelph.ca
with
your
Teams
ID/email
address
to
attend
the
seminars
online.
All
are
welcome
to
attend!
ABSTRACT: Cellular activities reply on proper functioning of a myriad of large biomolecular complexes. To understand the mechanisms by which these molecular machines work, it is crucial to obtain structural information as well as detailed characterization of their conformational dynamics. Conventional solution nuclear magnetic resonance (NMR) spectroscopy is limited to biomolecules with an upper size limit of ~40kDa; yet, with the development of solution NMR methodology and isotope labeling schemes in the recent decades, we can tackle large biomolecular complexes of up to 1 MDamolecular weight, allowing investigation into challenging biological supramolecular machines. Here we will present examples of using methyl-TROSY NMR to uncover structural and mechanistic details of large biomolecular complexes. We will focus on our previous and on-going work on an AAA+ protein, p97, which is a highly conserved molecular machine involved in a number ofcellular functions and has emerged as a promising target for cancer therapy. We have revealed cooperative conformational interconversion of its N-terminal domain and the implication in cofactor binding. We also characterized the complex between p97 and its adaptor protein p47, the complex of which plays an indispensable role in Golgi membrane fusion after mitosis. In addition, we will present our current efforts towards further defining the assembling and subunit exchange process of the p97 complex, as well as probing the mechanisms of two other AAA+ machines, human mitochondrial AAA proteases and bacterial DnaA.