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Tuberculosis serine hydrolase variable expression, isolation, and characterization under hypoxia conditions
Author(s) -
Severinac Luke Edward,
Johnson R. Jeremy
Publication year - 2018
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2018.32.1_supplement.796.20
Subject(s) - serine hydrolase , hydrolase , serine , mycobacterium tuberculosis , mycobacterium smegmatis , enzyme , biochemistry , chemistry , biology , tuberculosis , medicine , pathology
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 Information This 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 .