Biomedical subjects

Gene A Scarborough

Publications and source records attributed to Gene A Scarborough.

4 recordsLinked to original sources

PubMed2003

Rethinking the P-type ATPase problem.

There are very good reasons to stop thinking about the molecular mechanism of the P-type ion-translocating ATPases in terms of the traditional E1E2 model and to start thinking about it in more progressive ways. This makes it possible to see the ion-transport cycle as a rational series of discrete steps with well defined driving forces, including the crucial energy transduction step, where the chemical energy of ATP hydrolysis is exchanged for the osmotic energy of an ion gradient. Importantly, although major enzyme conformational changes accompany each of these steps, none of them drive the energy coupling reaction. Thus, neither the E1E2 model nor conformational energy coupling, the cornerstones of traditional thinking about the P-type ATPases, are reliable paradigms for future efforts to understand how these transporters work. Alternatives must be seriously considered.

Adenosine Triphosphate↗

PubMed2003

Why we must move on from the E1E2 model for the reaction cycle of the P-type ATPases.

Recent progress regarding the structure of the Ca(2+)-translocating ATPase of sarcoplasmic reticulum in several conformational states, and a substantial accumulation of biochemical information about this and other P-type ATPases, have put everything in place for the final convergence of biochemistry and structure that will lead to a complete understanding of the molecular mechanism of these membrane transport enzymes. But the common paradigm used to describe the reaction cycle of the P-type ATPases, the E1E2 model, is seriously flawed, and this is hindering our progress toward this goal. In this paper, it is first shown why the E1E2 model must be discarded. This is followed by a description of the P-type ATPase catalytic cycle that is much more consistent with the structural and biochemical information now available for these enzymes, and also brings to light the origin of the forces that drive the key reaction in the active transport cycle where high-affinity ion-binding sites are converted to low-affinity binding sites capable of releasing the transported ions against a considerable concentration gradient. This new model will therefore serve us better as we seek to unravel the final details of the molecular mechanism of active ion transport catalyzed by these enzymes. It is thus time to move on from the traditional E1E2 model.

Adenosine Triphosphatases↗

PubMed2002

Domain movements of plasma membrane H(+)-ATPase: 3D structures of two states by electron cryo-microscopy.

H(+)-ATPase is a P-type ATPase that transports protons across membranes using the energy from ATP hydrolysis. This hydrolysis is coupled to a conformational change between states of the protein, in which the proton-binding site is alternately accessible to the two sides of the membrane with an altered affinity. When isolated from Neurospora crassa, H(+)-ATPase is a 600 kDa hexamer of identical 100 kDa polypeptides. We have obtained the three-dimensional structures of both ligand-free and Mg(2+)/ADP-bound states of this complex using single-particle electron cryo- microscopy. Structural comparisons, together with the difference map, indicate that there is a rearrangement of the cytoplasmic domain on Mg(2+)/ADP binding, which consists of a movement of mass towards the 6-fold axis causing the structure to become more compact, accompanied by a modest conformational change in the transmembrane domain.

Adenosine Diphosphate↗

PubMed2002

Molecular mechanism of the P-type ATPases.

The recent determination of the structure of the Ca2+-ATPase of sarcoplasmic reticulum to atomic resolution in the Ca2+-bound state and to near atomic resolution in the Ca2+-free, decavanadate-bound state has paved the way for an ultimate complete understanding of the molecular mechanism of the P-type ATPases. Analysis of this new structure information together with the large amount of biochemical information about these enzymes that preceded it has produced important new revelations about how the P-type ATPases work. Most models propose that these transporters operate by a strictly conformational energy coupling mechanism in which conformational changes in the large cytoplasmic head region mechanically drive the ions to be transported from their binding sites in the transmembrane helix region 50 A away. However, while these enzymes do indeed undergo profound conformational changes, the available evidence suggests that they do not mechanically transduce the chemical energy of ATP hydrolysis into transmembrane ion gradients via these conformational changes. As an alternative, it is proposed that the effects of the chemical events that occur at the phosphorylation/dephosphorylation site in the cytoplasmic region are exerted on the ion-binding sites via two well-defined charge transfer pathways that electronically connect the chemical reaction site with the site of ion binding. The recognition of these charge transfer pathways provides rational explanations of all of the key biochemical features of the P-type ATPase catalytic cycle. Thus, although a few details await elucidation, a nearly complete understanding of the P-type ATPase reaction mechanism may be at hand.

Adenosine Triphosphatases↗