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Content archived on 2024-05-14

Structure-function relationship in plant plasma membrane proton pumps

Objective



Plant plasma membrane proton pumps (H+-ATPases) have a primary role in the coupling of metabolic energy to solute transport.
The objective of the consortium is the understanding of how the structure of these pumps is related to their function.
Plasma membrane H+-ATPases are not related to intracellular H+-ATPases such as FoFl type ATPases and vacuolar ATPases. They belong to the P-type ATPase family which among others includes the plasma membrane Na+/K+-, H+/K+- and Ca2+-ATPases of animals. So far, no Ptype ATPase structure has been solved. Thus, structural information on any P-type ATPase will be a breakthrough in understanding how these cation pumps carry out transport.
It will be a major aim to produce 2D and 3D crystals of purified plant plasma membrane H+-ATPase (Task 1). The structure of a 113 amino acid peptide comprising the Cterminal regulatory domain of the H+-ATPase will be solved separately by X-ray diffraction analysis of 3D crystals (Task 2). The information gained from obtaining the structure will be complemented by a search for individual amino acids important for H+-ATPase function (Task 3). The consortium brought together in this proposal is in a good position to realise these tasks because 1) an expression system allowing production of a single, homogenous and functional plant H+-ATPase has already been developed; 2) an in-vivo selection system for improved H+-ATPase activity is available; 3) the problem of structure determination of membrane proteins is circumvented by combining 2D and 3D crystallization, and 4) several P-type ATPases have already yielded 2D crystals, one of them by one of the partners.

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Coordinator

THE ROYAL VETERINARY AND AGRICULTURAL UNIVERSITY
EU contribution
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Address
10,Thorvaldsensvej 40
1870 FREDERISKSBERG
Denmark

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Participants (4)

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