| Autor: |
Erik Klontz, Juliet O. Obi, Yajing Wang, Gabrielle Glendening, Jahid Carr, Constantine Tsibouris, Sahthi Buddula, Shreeram Nallar, Alexei S. Soares, Dorothy Beckett, Jasmina S. Redzic, Elan Eisenmesser, Cheyenne Palm, Katrina Schmidt, Alexis H. Scudder, Trinity Obiorah, Kow Essuman, Jeffrey Milbrandt, Aaron Diantonio, Krishanu Ray, Daniel Deredge, M LD. Snyder, Greg A. Snyder |
| Rok vydání: |
2023 |
| Popis: |
Toll-like and Interleukin-1/18 receptor resistance (TIR) domain-containing proteins function as important signaling and immune regulatory molecules. TIR domain-containing proteins identified in eukaryotic and prokaryotic species also exhibit NAD+ hydrolase activity in select bacteria, plants, and mammalian cells. We report the crystal structure of theAcinetobacter baumanniiTIR domain protein (AbTir-TIR) with confirmed NAD+hydrolysis and map the conformational effects of its interaction with NAD+using HDX-MS. NAD+results in mild decreases in deuterium uptake at the dimeric interface. In addition, AbTir-TIR exhibits EX1 kinetics indicative of large cooperative conformational changes which are slowed down upon substrate binding. Additionally, we have developed label-free imaging using 2pFLIM which shows differences in bacteria expressing native and mutant NAD+ hydrolase-inactivated AbTir-TIREAprotein. Our observations are consistent with substrate-induced conformational changes reported in other TIR model systems with NAD+ hydrolase activity. These studies provide further insight into bacterial TIR protein mechanisms and their varying roles in biology. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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