Hr. Kaback, A MOLECULAR MECHANISM FOR ENERGY COUPLING IN A MEMBRANE-TRANSPORT PROTEIN, THE LACTOSE PERMEASE OF ESCHERICHIA-COLI, Proceedings of the National Academy of Sciences of the United Statesof America, 94(11), 1997, pp. 5539-5543
A mechanism for the coupled translocation of substrate and H+ by the l
actose permease of Escherichia coli is proposed, based on a variety of
experimental observations, The permease is composed of 12 alpha-helic
al rods that traverse the membrane with the N and C termini on the cyt
oplasmic face. Four residues are irreplaceable with respect to couplin
g, and the residues are paired-Arg-302 (helix IX) with Glu-325 (helix
X) and His-322 (helix X) with Glu-269 (helix VIII), In an adjacent reg
ion of the molecule at the interface between helices VIII and V is the
substrate translocation pathway, Because of this arrangement, interfa
cial changes between helices VIII and V are transmitted to the interfa
ce between helices IX and X and vice versa, Upon ligand binding, a str
uctural change at the interface between helices V and VIII disrupts th
e interaction between Glu-269 and His-322, Glu-269 displaces Glu-325 f
rom Arg-302, and Glu-325 is protonated. Simultaneously, protonated Glu
-325 becomes inaccessible to water, which drastically increases its pK
(a), In this configuration, the permease undergoes a freely reversible
conformational change that corresponds to translocation of the ternar
y complex, To return to ground state after release of substrate, the A
rg-302-Glu-325 interaction must be reestablished, which necessitates l
oss of H+ from Glu-325, The H+ is released into a water-filled crevice
between helices IX and X which becomes transiently accessible to both
sides of the membrane due to a change in helix tilt, where it is acte
d upon equally by either the membrane potential or the pH gradient acr
oss the membrane.