"Single-molecule protein translocation through nano pores"Who: Dr. David Rodriguez-Larrea, Dept. of Chemistry, University of Oxford Place: nanoGUNE seminar room, Tolosa Hiribidea 76, Donostia - San Sebastian Date: Monday, 24 March 2014, 11:00 Nanopores have emerged as a powerful tool for the analysis of single
molecules in solution. The ionic current passed through them is disrupted by
analytes in a characteristic and measurable way. Protein nanopores offer high reproducibility
in pore dimensions and the ability to introduce site-specific chemical
modifications. This has led to the use of these systems as a platform for the
study of chemical reactions at the single-molecule level and as an approach
towards ultra-fast and cheap sequencing of DNA. Here I will describe a nanopore system that allows tracking protein
co-translocational unfolding at the single-molecule level. Several biological
processes require protein translocation through narrow pores. For example, the
traffic of proteins across membranes is frequently mediated by them. Furthermore,
the proteasome along with chaperones from the AAA+ family unfold proteins by pulling
them through a narrow pore. Our single-molecule measurements give insight on
how proteins unfold through pores and refold after translocation. Protein function, on the other hand, is often regulated by chemical
modifications of specific residues (i.e. post-translational modifications).
Based on the ability of nanopores to detect sequence features I will show that
single-molecule and site-specific detection of post-translational modifications
is feasible. |