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Master of Science (MS)
Anthony-Cahill, Spencer J.
Spiegel, P. Clint
Smirnov, Serge L.
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 collect spectra with resolved, well defined peaks that can be assigned, thereby providing valuable structural information. We have assigned 89.2% of backbone atoms and 87.4% of sidechain atoms. Assignment of Tae1 was performed with 15N-HSQC, HNCA, HNCOCA, HNCACB, CBCACONH, HCCH COSY, HCCH TOCSY, and HCONH TOCSY experiments. Peptidoglycan binding experiments were performed using via 15N-HSQC to monitor backbone residues and 13C-HSQC to monitor sidechain residues. So far, these experiments have not revealed the molecular mechanism by which Tae1 recognizes its specific substrate; however, with the very high degree of assignment achieved in NMR experimentation, once a minimal binding fragment has been isolated determination of the binding mechanism will be easily achieved.
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Henderson, Robert C. (Robert Corey), "Mapping Interactions Between the Type-VI Secretion System Effector tAE1 and Its Putative Substrates Using NMR Spectroscopy" (2016). WWU Masters Thesis Collection. 521.