Gas tunnel engineering of prolyl hydroxylase reprograms hypoxia signaling in cells.

ABSTRACT: Cells have evolved intricate mechanisms for recognizing and responding to changes in oxygen (O₂) concentrations. Here, we have reprogrammed cellular hypoxia (low O₂) signaling via gas tunnel engineering of prolyl hydroxylase 2 (PHD2), a non-heme iron dependent O₂ sensor. Using computational modeling and protein engineering techniques, we identify a gas tunnel and critical residues therein that limit the flow of O₂ to PHD2's catalytic core. We show that systematic modification of these residues can open the constriction topology of PHD2's gas tunnel. Using kinetic stopped-flow measurements with NO as a surrogate diatomic gas, we demonstrate up to 3.5-fold enhancement in its association rate to the iron center of tunnel-engineered mutants. Our most

SUBMITTER: Windsor P 

PROVIDER: S-EPMC10441328 | biostudies-literature | 2024 May

REPOSITORIES: biostudies-literature