Sucrose- and H-dependent charge movements associated with the gating of sucrose transporter ZmSUT1.
BACKGROUND
In contrast to man the majority of higher plants use sucrose as mobile carbohydrate. Accordingly proton-driven sucrose transporters are crucial for cell-to-cell and long-distance distribution within the plant body. Generally very negative plant membrane potentials and the ability to accumulate sucrose quantities of more than 1 M document that plants must have evolved transporters with unique structural and functional features.


RESULTS
To unravel the functional properties of one specific high capacity plasma membrane sucrose transporter in detail, we expressed the sucrose/H(+) co-transporter from maize ZmSUT1 in Xenopus oocytes. Application of sucrose in an acidic pH environment elicited inward proton currents. Interestingly the sucrose-dependent H(+) transport was associated with a decrease in membrane capacitance (C(m)). In addition to sucrose C(m) was modulated by the membrane potential and external protons. In order to explore the molecular mechanism underlying these C(m) changes, presteady-state currents (I(pre)) of ZmSUT1 transport were analyzed. Decay of I(pre) could be best fitted by double exponentials. When plotted against the voltage the charge Q, associated to I(pre), was dependent on sucrose and protons. The mathematical derivative of the charge Q versus voltage was well in line with the observed C(m) changes. Based on these parameters a turnover rate of 500 molecules sucrose/s was calculated. In contrast to gating currents of voltage dependent-potassium channels the analysis of ZmSUT1-derived presteady-state currents in the absence of sucrose (I =  Q/τ) was sufficient to predict ZmSUT1 transport-associated currents.


CONCLUSIONS
Taken together our results indicate that in the absence of sucrose, 'trapped' protons move back and forth between an outer and an inner site within the transmembrane domains of ZmSUT1. This movement of protons in the electric field of the membrane gives rise to the presteady-state currents and in turn to C(m) changes. Upon application of external sucrose, protons can pass the membrane turning presteady-state into transport currents.
Carpaneto
Armando
A
Istituto di Biofisica-CNR, Genova, Italy.
Koepsell
Hermann
H
Bamberg
Ernst
E
Hedrich
Rainer
R
Geiger
Dietmar
D
D000818:Animals; D001692:Biological Transport; D026901:Membrane Transport Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D009865:Oocytes / Q000378:metabolism; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D011522:Protons; D013395:Sucrose / Q000378:metabolism*; D014982:Xenopus laevis; D003313:Zea mays / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D026901:Membrane Transport Proteins; D010940:Plant Proteins; D011522:Protons; D013395:Sucrose