Different inhibitory capacities of human and mouse KLRG1 are linked to distinct disulfide-mediated oligomerizations

The killer cell lectin-like receptor G1 (KLRG1) is a cadherin-binding inhibitory receptor expressed by NK and T cells in humans and mice. Although structural and ligand-binding properties of human (h) and mouse (m) KLRG1 are very similar, KLRG1-mediated inhibition under physiological conditions is only observed with human lymphocytes. Using a well-defined in vitro system, we demonstrate here that mKLRG1 exhibits a significantly lower inhibitory capacity compared with the human homolog. Biochemical analyses further showed that mKLRG1 formed monomers and disulfide-linked dimers, trimers, and tetramers whereas hKLRG1 was exclusively present as disulfide-linked dimer. Mutational analysis revealed a crucial role of Cys(62) present in the stalk region of mKLRG1 but not of hKLRG1 for oligomer formation. Strikingly, mimicking hKLRG1 by replacement of Cys(62) in mKLRG1 by glutamine prevented tri- and tetramer formation and increased the inhibitory capacity. Furthermore, mutated mKLRG1 molecules that were unable to form disulfide-linked dimers at all or at a decreased level lacked inhibitory activity. These data indicate that only dimeric KLRG1 entities exhibit potent inhibitory capacities. The lower inhibitory capacity of mKLRG1 compared with hKLRG1 can thus be rationalized by a decreased proportion of dimeric entities, which can be pinpointed to a single amino acid.