Enantiomeric oxaziridine probes systematically reveal that privileged methionine residues undergo stereospecific oxidation to (S)- or (R)-methionine sulfoxide across proteomes, with structural preferences for alpha helices versus beta sheets respectively, and demonstrate that (R)-methionine sulfoxide formation at BPHL M69 allosterically inhibits enzyme activity through an MsrB2-regulated pathway that controls cellular protein N-homocysteinylation levels and downstream metabolic processes.

Life is predicated on chirality, a molecular asymmetry akin to the left and right versions of human hands. Here we show that privileged protein residues are predisposed for chiral regulation. We developed enantiomeric oxaziridine reagents that systematically identify pro-(S) and pro-(R) methionine oxidation sites across proteomes that can be erased by stereospecific methionine sulfoxide reductase enzymes A and B, respectively. These probes reveal that chiral regulation of methionine oxidation-reduction processes can allosterically regulate protein function, as shown in cell and murine models of oxidative stress where selective (R)-methionine sulfoxide formation on M69 of biphenyl hydrolase-like protein leads to hydrolase inhibition and amplification of proteome N-homocysteinylation modifications. This work introduces a platform for characterizing sites of asymmetric methionine oxidation and the functional consequences concomitant with an individual chiral single-atom modification.