Building the stator of the yeast vacuolar-ATPase: Specific interaction between subunits E and G
Publikation: Beitrag in Fachzeitschrift › Forschungsartikel › Beigetragen › Begutachtung
Beitragende
Abstract
The vacuolar (H+)-ATPase (or V-ATPase) is a membrane protein complex that is structurally related to F1 and F0 ATP syntheses. The V-ATPase is composed of an integral domain (V0) and a peripheral domain (V1) connected by a central stalk and up to three peripheral stalks. The number of peripheral stalks and the proteins that comprise them remain controversial. We have expressed subunits E and G in Escherichia coli as maltose binding protein fusion proteins and detected a specific interaction between these two subunits. This interaction was specific for subunits E and G and was confirmed by co-expression of the subunits from a bicistronic vector. The EG complex was characterized using size exclusion chromatography, cross-linking with short length chemical cross-linkers, circular dichroism spectroscopy, and electron microscopy. The results indicate a tight interaction between subunits E and G and revealed interacting helices in the EG complex with a length of about 220 Å. We propose that the V-ATPase EG complex forms one of the peripheral stators similar to the one formed by the two copies of subunit b in F-ATPase.
Details
Originalsprache | Englisch |
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Seiten (von - bis) | 40670-40676 |
Seitenumfang | 7 |
Fachzeitschrift | Journal of Biological Chemistry |
Jahrgang | 279 |
Ausgabenummer | 39 |
Publikationsstatus | Veröffentlicht - 24 Sept. 2004 |
Peer-Review-Status | Ja |
Extern publiziert | Ja |
Externe IDs
PubMed | 15292229 |
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ORCID | /0000-0002-4482-6010/work/142251041 |