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Abstract

The C-type lectin receptor langerin plays a vital role in the mammalian defense against invading pathogens. Langerin requires a ²⁺+ co-factor, the binding affinity of which is regulated by pH. Thus, ²⁺+ is bound when langerin is on the membrane, but released when langerin and its pathogen substrate traffic to the acidic endosome, allowing the substrate to be degraded. The change in pH is sensed by protonation of the allosteric pH-sensor histidine H294. However, the mechanism by which ²⁺+ is released from the buried binding site is not clear. We studied the structural consequences of protonating H294 by molecular dynamics simulations (total simulation time: about 120 μs) and Markov models. We discovered a relay mechanism in which a proton is moved into the vicinity of the ²⁺+-binding site without transferring the initial proton from H294. Protonation of H294 unlocks a conformation in which a protonated lysine side-chain forms a hydrogen bond with a ²⁺+-coordinating aspartic acid. This destabilizes ²⁺+ in the binding pocket, which we probed by steered molecular dynamics. After ²⁺+-release, the proton is likely transferred to the aspartic acid and stabilized by a dyad with a nearby glutamic acid, triggering a conformational transition and thus preventing ²⁺+-rebinding. These results show how pH-regulation of a buried orthosteric binding site from a solvent-exposed allosteric pH-sensor can be realized by information transfer through a specific chain of conformational arrangements.