Abstract
Tuberculosis (TB), caused by infection by Mycobacterium tuberculosis, is the deadliest infectious disease worldwide. Complicating the treatment of TB is its ability to transition between an intransient dormant state and an active infectious state. One class of enzymes required for the maintenance of this dormant state and the transition into the active state is serine hydrolases. Our research goal was to globally characterize serine hydrolase activity in actively growing M. smegmatis and under hypoxia conditions that mimic dormant growth. To measure hydrolase activity, lysates were prepared under different growth conditions and then separated by native‐PAGE. Native‐PAGE gels were incubated with a library of fluorogenic ester substrates and hydrolase activity quantitated based on fluorogenic substrate activation. By following the shift in fluorogenic activation, we were able to measure changes in hydrolase protein expression versus fluorogenic substrate and in relation to induction of dormant growth conditions. Reactive hydrolases were excised from the gel and identified by mass spectrometry. Our results provided a global map of mycobacterial serine hydrolase activity and identified starting points for finding novel drug targets for dormant TB.Support or Funding InformationThis work was supported by a grant from the National Institutes of Health (NIH 1 R15 GM110641‐01A1)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.