Proton Transfer Limits Protein Translocation Rate by the Thylakoid ΔpH/Tat Machinery

Abstract

The thylakoid transmembrane DeltapH is the sole energy source driving translocation of precursor proteins by the DeltapH/Tat machinery. Consequently, proton translocation must be coupled to precursor translocation. For the precursor of the 17 kDa protein of the oxygen-evolving complex (pOE17), the protein translocation process is characterized by a steep drop in efficiency at an external pH below 7.0 and above 8.7. As the membrane DeltapH is virtually unaffected from pH 6.5 to 9.2, the loss in import efficiency is a consequence of the titration of multiple residues within the translocation machinery. Transport is retarded by a factor of 2-3 in deuterium oxide (D(2)O) relative to water, strongly suggesting that proton-transfer reactions limit translocation rate. The solvent isotope effect manifests itself after the precursor binds to the membrane, indicating that the rate-limiting step is a later event in the transport process.