cb9b00788_si_002.xlsx (9.87 kB)
Discovering the Microbial Enzymes Driving Drug Toxicity with Activity-Based Protein Profiling
dataset
posted on 2019-12-12, 20:44 authored by Parth
B. Jariwala, Samuel J. Pellock, Dennis Goldfarb, Erica W. Cloer, Marta Artola, Joshua B. Simpson, Aadra P. Bhatt, William G. Walton, Lee R. Roberts, Michael B. Major, Gideon J. Davies, Herman S. Overkleeft, Matthew R. RedinboIt is increasingly clear that interindividual variability
in human
gut microbial composition contributes to differential drug responses.
For example, gastrointestinal (GI) toxicity is not observed in all
patients treated with the anticancer drug irinotecan, and it has been
suggested that this variability is a result of differences in the
types and levels of gut bacterial β-glucuronidases (GUSs). GUS
enzymes promote drug toxicity by hydrolyzing the inactive drug–glucuronide
conjugate back to the active drug, which damages the GI epithelium.
Proteomics-based identification of the exact GUS enzymes responsible
for drug reactivation from the complexity of the human microbiota
has not been accomplished, however. Here, we discover the specific
bacterial GUS enzymes that generate SN-38, the active and toxic metabolite
of irinotecan, from human fecal samples using a unique activity-based
protein profiling (ABPP) platform. We identify and quantify gut bacterial
GUS enzymes from human feces with an ABPP-enabled proteomics pipeline
and then integrate this information with ex vivo kinetics
to pinpoint the specific GUS enzymes responsible for SN-38 reactivation.
Furthermore, the same approach also reveals the molecular basis for
differential gut bacterial GUS inhibition observed between human fecal
samples. Taken together, this work provides an unprecedented technical
and bioinformatics pipeline to discover the microbial enzymes responsible
for specific reactions from the complexity of human feces. Identifying
such microbial enzymes may lead to precision biomarkers and novel
drug targets to advance the promise of personalized medicine.