Open AccessReactivity of N-acyl hydrazone probes with the mammalian proteome
Author(s) -
Tyler A. Shaw,
Megan H. Powdrill,
Allison R. Sherratt,
Keira Garland,
Bin-Jie Li,
André M. Beauchemin,
John Paul Pezacki
Publication year - 2021
Publication title -
rsc medicinal chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.754
H-Index - 55
ISSN - 2632-8682
DOI - 10.1039/d1md00027f
Subject(s) - hydrazone , chemistry , proteome , cytidine deaminase , cysteine , serine , covalent bond , biochemistry , enzyme , active site , proteomics , serine hydrolase , combinatorial chemistry , stereochemistry , organic chemistry , gene
Small molecule probes with distinct reactivities are useful tools for the identification and characterization of protein modifications and function. Herein, we show that hydrazone probes with an N -carbamate structural motif react differently from N -carbamates within the human proteome. Mass spectrometry analysis of probe-treated mammalian cell lysates identified several proteins that were covalently modified by the hydrazone probes, including the cytidine deaminase APOBEC3A. We used this enzyme as a model to explore the reactivity of the probes with amino acid residues using LC-MS/MS. Both reactive serine and cysteine residues outside of the enzyme active site were covalently modified. A 1-napthol leaving group provided the most extensive reactivity. These results confirm a unique chemotype for hydrazone probes which can be further optimized to target distinct targets of the human proteome.