Probe for selectively characterizing enzymes involved in xenobiotic metabolism and method of making and using the same
AssigneeBattelle Memorial Institute
InventorsAaron T. Wright, Susan Ramos-Hunter, Christopher Whidbey
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Abstract
Activity-based probes that can be used to selectively identify and characterize enzymes that are involved in different phases of xenobiotic metabolism in a host and its microbiota population(s) are described. The activity-based probes described specifically label only their target active enzymes involved in xenobiotic metabolism and therefore provide a measurement of true protein functional activity rather than transcript or protein abundance. The activity-based probes also provide multimodal profiling of these active enzymes. Methods for preparing the activity based probes and exemplary methods for their use also are disclosed.
Claims22
A probe having a structure satisfying Formula II
- wherein
- ERG, if present, is
- S(O)2OH, or an anionic form thereof;
- P(O(OH)2, or an anionic form thereof;
- a halogen;
- or —OPh-CH2—ONO2;
- EBG has a structure satisfying a formula selected from Formulas IIAEBG-IIJEBG
- wherein Y is CH3 or CF3;
- Y′ is O, NO2, or —N═NR″, wherein R″ is a dye or other reporting moiety;
- and m′ is an integer selected from 0 to 5;
- and R′ is selected from aldehyde, ketone, ester, carboxylic acid, acyl, acyl halide, cyano, sulfonate, nitro, nitroso, quaternary amine, CF3, alkyl halide, or combinations thereof;
- Linkera comprises an aliphatic group, a heteroaliphatic group, an aromatic group, an aliphatic-aromatic group, a heteroaliphatic-aromatic group, a heteroaromatic group, an aliphatic-heteroaromatic group, or a heteroaliphatic-heteroaromatic group;
- each of Linkerb, and Linkerc, if present, independently comprises an aliphatic group, a heteroaliphatic group, an aromatic group, an aliphatic-aromatic group, a heteroaliphatic-aromatic group, a heteroaromatic group, an aliphatic-heteroaromatic group, or a heteroaliphatic-heteroaromatic group;
- R, if present, is a group comprising a carbamate or a carbonate;
- and
- each of n, m, p, and q independently is 0 or 1;
- provided that one of the following applies:
- (a) one or more Tags are present, and each Tag is independently a fluorophore, a binding partner of an affinity-based binding pair, a quantum dot, or a dye;
- or
- (b) one or more pTags are present and each pTag is independently an azide or an alkyne.
A probe selected from:
A probe having a structure satisfying Formula II
- wherein
- ERG is
- EBG has a structure satisfying a formula selected from
- wherein Y′ is O;
- m′ is an integer selected from 0 to 5;
- and each occurrence of R′ is independently selected from aldehyde, ketone, ester, carboxylic acid, acyl, acyl halide, cyano, sulfonate, nitro, nitroso, quaternary amine, CF3, alkyl halide, or combinations thereof;
- Linkera comprises an aliphatic group, a heteroaliphatic group, an aromatic group, an aliphatic-aromatic group, a heteroaliphatic-aromatic group, a heteroaromatic group, an aliphatic-heteroaromatic group, or a heteroaliphatic-heteroaromatic group;
- each of Linkerb, and Linkerc, if present, independently comprises an aliphatic group, a heteroaliphatic group, an aromatic group, an aliphatic-aromatic group, a heteroaliphatic-aromatic group, a heteroaromatic group, an aliphatic-heteroaromatic group, or a heteroaliphatic-heteroaromatic group;
- R, if present, is a group comprising a carbamate or a carbonate;
- each of n, m, p, and q independently is 0 or 1;
- and
- provided that one of the following applies:
- (a) one or more Tags are present, and each Tag is independently a fluorophore, a binding partner of an affinity-based binding pair, a quantum dot, or a dye;
- or
- (b) one or more pTags are present, and each pTag is independently an azide or an alkyne.
A probe selected from:
Dependencies are inferred from the claim text.