Characterization of a Key Mycobacterium tuberculosis Lipase, LipY Public Deposited

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  • March 19, 2019
Creator
  • Garrett, Christopher
    • Affiliation: School of Medicine, Department of Biochemistry and Biophysics
Abstract
  • The causative agent of tuberculosis, Mycobacterium tuberculosis is estimated to be present in roughly a third of the world’s population. One of the hallmarks of the pathogen is its reliance on lipids as a source of energy in order to survive in a non-replicating state and ultimately cause infection. A key player in lipid utilization by M. tuberculosis is the triglyceride lipase, LipY. LipY is the rate-limiting catalyst in the mobilization of free fatty acids from triglycerides acting on lipid bodies both within the tubercule bacillus and outside the bacterium. LipY also belongs to the PE/PPE family of proteins, a group of approximately 169 virulence-associated proteins characterized by a proline-glutamate or proline-proline-glutamate motif at their N-termini. It has been suggested that the PE domain of LipY may act as a regulator of enzymatic activity. Using several biochemical and activity-based approaches, we established a method for purifying LipY with improved resolution. We also introduced a rapid and reliable method for quantification of active sites for serine hydrolases using activity-based protein profiling (ABPP). Finally, by imploring these methodologies, we discovered that the PE domain of LipY acts as a non-competitive inhibitor of LipY enzymatic activity when it is attached to the mature lipase. The methodologies and evidence presented here may lead to discovery of additional PE/PPE family enzymes with roles in M. tuberculosis lipid metabolism and/or virulence. This work may also lead to expansion of our understanding of the roles of PE/PPE proteins and domains.
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  • In Copyright
Advisor
  • Braunstein, Miriam
  • Redinbo, Matthew R.
  • Holly, Stephen
  • Neher, Saskia
  • Carter, Charles
Degree
  • Doctor of Philosophy
Degree granting institution
  • University of North Carolina at Chapel Hill Graduate School
Graduation year
  • 2015
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  • Chapel Hill, NC
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