Multifunctional Activity-Based Protein Profiling of the Developing Lung

Lung diseases and disorders are a
leading cause of death among
infants. Many of these diseases and disorders are caused by premature
birth and underdeveloped lungs. In addition to developmentally related
disorders, the lungs are exposed to a variety of environmental contaminants
and xenobiotics upon birth that can cause breathing issues and are
progenitors of cancer. In order to gain a deeper understanding of
the developing lung, we applied an activity-based chemoproteomics
approach for the functional characterization of the xenometabolizing
cytochrome P450 enzymes, active ATP and nucleotide binding enzymes,
and serine hydrolases using a suite of activity-based probes (ABPs).
We detected P450 activity primarily in the postnatal lung; using our
ATP-ABP, we characterized a wide range of ATPases and other active
nucleotide- and nucleic acid-binding enzymes involved in multiple
facets of cellular metabolism throughout development. ATP-ABP targets
include kinases, phosphatases, NAD- and FAD-dependent enzymes, RNA/DNA
helicases, and others. The serine hydrolase-targeting probe detected
changes in the activities of several proteases during the course of
lung development, yielding insights into protein turnover at different
stages of development. Select activity-based probe targets were then
correlated with RNA in situ hybridization analyses of lung tissue
sections.