Mining the cellular inventory of pyridoxal phosphate-dependent enzymes with functionalized cofactor mimics.

ABSTRACT: Pyridoxal phosphate (PLP) is an enzyme cofactor required for the chemical transformation of biological amines in many central cellular processes. PLP-dependent enzymes (PLP-DEs) are ubiquitous and evolutionarily diverse, making their classification based on sequence homology challenging. Here we present a chemical proteomic method for reporting on PLP-DEs using functionalized cofactor probes. We synthesized pyridoxal analogues modified at the 2'-position, which are taken up by cells and metabolized in situ. These pyridoxal analogues are phosphorylated to functional cofactor surrogates by cellular pyridoxal kinases and bind to PLP-DEs via an aldimine bond which can be rendered irreversible by NaBH₄ reduction. Conjugation to a reporter tag enables the subsequent identification of

SUBMITTER: Hoegl A 

PROVIDER: S-EPMC6252082 | biostudies-literature | 2018 Dec

REPOSITORIES: biostudies-literature