Small-molecules that covalently react with a human prolyl hydroxylase - towards activity modulation and substrate capture.

Jacob T Bush,Tzu-Lan Yeh,Jasmin Mecinović,Anthony Tumber,Roman Belle,Emily Flashman,Holger Kramer,Rasheduzzaman Chowdhury,Christopher J Schofield,Robert K Leśniak

doi:10.1039/c8cc07706a

Abstract

We describe covalently binding modulators of the activity of human prolyl hydroxylase domain 2 (PHD2) and studies towards a strategy for photocapture of PHD2 substrates. Reversible active site binding of electrophile bearing compounds enables susbsequent covalent reaction with a lysine residue (K408) in the flexible C-terminal region of PHD2 to give a modified protein that retains catalytic activity.

Highlights

Schofield et al Small-molecules that covalently react with a human prolyl hydroxylase – towards activity modulation and substrate capture
We describe covalently binding modulators of the activity of human prolyl hydroxylase domain 2 (PHD2) and studies towards a strategy for photocapture of PHD2 substrates
PHD2 structures suggest that its active site does not contain Lys- or Cys-residues; Lys-residues are present in a loop (K246, K251) and the C-terminal section (K400, K402, K408, K416, K423), both of which are flexible and associated with the active site entrance (Fig. 1A).[14,15]

Summary

Introduction

Schofield et al Small-molecules that covalently react with a human prolyl hydroxylase – towards activity modulation and substrate capture Reversible active site binding of electrophile bearing compounds enables susbsequent covalent reaction with a lysine residue (K408) in the flexible C-terminal region of PHD2 to give a modified protein that retains catalytic activity. Analysis of 1 after 5 min indicated complete condensation; with 2, reaction was slower, with an apparent non-covalent adduct predominating after 5 min, with B30% condensation (Fig. S2, ESI†).

Results

Conclusion