Time-resolved NMR detection of prolyl-hydroxylation in intrinsically disordered region of HIF-1α.

ABSTRACT: Prolyl-hydroxylation is an oxygen-dependent posttranslational modification (PTM) that is known to regulate fibril formation of collagenous proteins and modulate cellular expression of hypoxia-inducible factor (HIF) α subunits. However, our understanding of this important but relatively rare PTM has remained incomplete due to the lack of biophysical methodologies that can directly measure multiple prolyl-hydroxylation events within intrinsically disordered proteins. Here, we describe a real-time ¹³C-direct detection NMR-based assay for studying the hydroxylation of two evolutionarily conserved prolines (P402 and P564) simultaneously in the intrinsically disordered oxygen-dependent degradation domain of hypoxic-inducible factor 1α by exploiting the "proton-less" nature of prolines

SUBMITTER: He W 

PROVIDER: S-EPMC11406255 | biostudies-literature | 2024 Sep

REPOSITORIES: biostudies-literature