An activity-dependent proximity ligation platform for spatially resolved quantification of active enzymes in single cells

Gang Li, Jeffrey E. Montgomery, Mark A. Eckert, Jae Won Chang, Samantha M. Tienda, Ernst Lengyel and Raymond E. Moellering ()
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Gang Li: The University of Chicago
Jeffrey E. Montgomery: The University of Chicago
Mark A. Eckert: The University of Chicago
Jae Won Chang: The University of Chicago
Samantha M. Tienda: The University of Chicago
Ernst Lengyel: The University of Chicago
Raymond E. Moellering: The University of Chicago

Nature Communications, 2017, vol. 8, issue 1, 1-12

Abstract: Abstract Integration of chemical probes into proteomic workflows enables the interrogation of protein activity, rather than abundance. Current methods limit the biological contexts that can be addressed due to sample homogenization, signal-averaging, and bias toward abundant proteins. Here we report a platform that integrates family-wide chemical probes with proximity-dependent oligonucleotide amplification and imaging to quantify enzyme activity in native contexts with high spatial resolution. Application of this method, activity-dependent proximity ligation (ADPL), to serine hydrolase and cysteine protease enzymes enables quantification of differential enzyme activity resulting from endogenous changes in localization and expression. In a competitive format, small-molecule target engagement with endogenous proteins in live cells can be quantified. Finally, retention of sample architecture enables interrogation of complex environments such as cellular co-culture and patient samples. ADPL should be amenable to diverse probe and protein families to detect active enzymes at scale and resolution out of reach with current methods.

Date: 2017
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DOI: 10.1038/s41467-017-01854-0

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