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Crystallization, structure and dynamics of the proton-translocating P-type ATPase
1Department of Pharmacology, University of North Carolina, School of Medicine, Chapel Hill, NC 27599, USA. gas@med.unc.edu
The Journal of Experimental Biology
|December 22, 1999
Summary
Researchers crystallized the Neurospora crassa H(+)-ATPase (H(+) P-ATPase) to reveal its structure. This study provides a model for the enzyme's transport cycle and driving forces.
Area of Science:
- Biochemistry
- Structural Biology
- Membrane Proteins
Background:
- The Neurospora crassa plasma membrane H(+)-ATPase (H(+) P-ATPase) is a crucial proton pump.
- Understanding its structure is key to elucidating its transport mechanism.
Purpose of the Study:
- To determine the structure of the H(+) P-ATPase complex.
- To provide insights into the enzyme's catalytic cycle and ion transport mechanism.
Main Methods:
- Crystallization of the H(+) P-ATPase complex using dodecylmaltoside and polyethylene glycol.
- Electron crystallographic analysis of two-dimensional crystals.
- Hydrogen/deuterium exchange experiments.
Main Results:
- Elucidation of the H(+) P-ATPase structure at 0.8 nm resolution in the membrane plane.
- Identification of ten transmembrane helices and distinct cytoplasmic domains.
- Evidence for rigid-body interdomain movements during conformational changes.
Conclusions:
- The crystal packing involves protein-protein and detergent-detergent interactions.
- A rational model for the H(+) P-ATPase transport cycle is proposed based on structure and dynamics.
- Atomic resolution is needed for a complete understanding of the ion transport mechanism.