The plasma membrane ATPase of Neurospora: a proton-pumping electroenzyme.
Probably the best marker enzyme for plasma membranes of eukaryotic cells is a magnesium-dependent, vanadate-inhibited ATPase whose primary function is the transmembrane transport of cations. In animal cells, different species of the enzyme transport different cations: sodium ions released in unequal exchange for potassium ions, calcium ions extruded alone (perhaps), or protons secreted in equal exchange for potassium ions. But in plants and fungi only proton secretion has been clearly demonstrated. A useful model cell for studying the proton-secreting ATPase has been the ascomycete fungus Neurospora, in which the enzyme drives an outward current of protons that can exceed 50 microA/cm2 and can support membrane potentials greater than 300 mV. Both thermodynamic and kinetic studies have shown that the proton-pumping ATPase of Neurospora normally transports only a single proton for each ATP molecule split; and kinetic modelling studies have suggested (contrary to conventional assumptions) that the fast steps in the overall reaction are transmembrane transit of the proton and its dissociation following transport, while the slow steps are the binding of protons and/or ATP. The primary structure of the Neurospora enzyme, recently deduced by gene sequencing, is very close to that of the yeast (Saccharomyces) enzyme, and the hydropathic patterns for both closely resemble those for the animal-cell plasma-membrane ATPases. All of these enzymes appear to have 6-10 membrane-spanning alpha-helices, plus a large cytoplasmic headgroup which bears the catalytic nucleotide-binding site. Structural data, taken together with the electrical-kinetic behavior, suggest that the catalytic headgroup functions as an energized gate for protons. From a geometric point of view, action of such a gate would transfer the membrane field across the "transported" ion, rather than vice versa.
D000251:Adenosine Triphosphatases / Q000378:metabolism*; D001692:Biological Transport; D002462:Cell Membrane / Q000201:enzymology / Q000502:physiology; D055598:Chemical Phenomena; D002621:Chemistry; D004594:Electrophysiology; D007700:Kinetics; D008954:Models, Biological; D009491:Neurospora / Q000201:enzymology* / Q000502:physiology; D011522:Protons
D011522:Protons; D000251:Adenosine Triphosphatases