Mapping interactions between the type-VI secretion system effector Tae1 and its putative substrates using NMR spectroscopy
Research Mentor(s)
Anthony-Cahill, Spencer J.
Description
Tae1 is an amidase produced by gram negative Pseudomonas bacteria that attacks the peptidoglycan layer in the cell walls of neighboring bacteria after secretion through the Type VI secretion system (T6S). The goal of our work is mapping interactions between the type-VI-secretion system effector Tae1 and its putative substrates using nuclear magnetic resonance (NMR) spectroscopy. Tae1 is amenable to NMR in that we are able to gather spectra with resolved, well defined peaks that can be assigned providing valuable structural information. We have been able to achieve nearly complete assignment of the backbone atoms, and roughly 85% coverage for the side-chain atoms of Tae1 via 15N-HSQC, HCCH COSY, HCCH TOCSY, and HCONH TOCSY experiments. Current work is focused on interpreting data from binding experiments of Tae1 with peptidoglycan via 13C-HSQC. From these binding experiments we hope to better understand the peptidoglycan cleavage specificity of Tae1.
Document Type
Event
Start Date
14-5-2015 10:00 AM
End Date
14-5-2015 2:00 PM
Department
Chemistry
Genre/Form
student projects; posters
Subjects – Topical (LCSH)
Amidases--Experiments; Peptidoglycans--Experiments; Pathogenic bacteria; Drug resistance in microorganisms; Nuclear magnetic resonance
Type
Image
Rights
Copying of this document in whole or in part is allowable only for scholarly purposes. It is understood, however, that any copying or publication of this documentation for commercial purposes, or for financial gain, shall not be allowed without the author's written permission.
Language
English
Format
application/pdf
Mapping interactions between the type-VI secretion system effector Tae1 and its putative substrates using NMR spectroscopy
Tae1 is an amidase produced by gram negative Pseudomonas bacteria that attacks the peptidoglycan layer in the cell walls of neighboring bacteria after secretion through the Type VI secretion system (T6S). The goal of our work is mapping interactions between the type-VI-secretion system effector Tae1 and its putative substrates using nuclear magnetic resonance (NMR) spectroscopy. Tae1 is amenable to NMR in that we are able to gather spectra with resolved, well defined peaks that can be assigned providing valuable structural information. We have been able to achieve nearly complete assignment of the backbone atoms, and roughly 85% coverage for the side-chain atoms of Tae1 via 15N-HSQC, HCCH COSY, HCCH TOCSY, and HCONH TOCSY experiments. Current work is focused on interpreting data from binding experiments of Tae1 with peptidoglycan via 13C-HSQC. From these binding experiments we hope to better understand the peptidoglycan cleavage specificity of Tae1.