Abstract
The isolated glycine betaine uptake carrier BetP from Corynebacterium glutamicum was reconstituted in Escherichia coli phospholipid liposomes and its response to osmotic stress studied. The transport activity of BetP, which was previously shown to comprise both osmosensory and osmoregulatory functions, was used to identify the nature of the physicochemical stimulus related to hyperosmotic stress. Putative factors modulating transport activity in response to osmotic stress were dissected. These include type, osmolality and concentration of solutes in the internal and/or external compartment (cationic, anionic, zwitterionic, neutral), as well as membrane strain as a response to increased osmolality. Osmoresponsive activation of BetP was independent of any external factor and of physical alterations of the membrane, but was triggered by a change in the internal K+ concentration. Activation did not depend on the type of anion present and was K+ (or Cs+ and Rb+) specific, as choline and NH(4)+ did not trigger BetP activity. The half-maximal activation of BetP in E.coli phospholipid liposomes was correlated to an internal concentration of 221 +/- 23 mM K+.
MeSH Terms
Bacterial Proteins
Betaine/metabolism
Carrier Proteins/metabolism
Cations, Monovalent/metabolism
Corynebacterium/metabolism
Cytoplasm/metabolism
Osmotic Pressure
Potassium/metabolism
Protein Conformation
Proteolipids
Signal Transduction
Symporters
Chemicals
Bacterial Proteins
BetP protein, Corynebacterium glutamicum
Carrier Proteins
Cations, Monovalent
Proteolipids
Symporters
proteoliposomes
Betaine
Potassium
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Rübenhagen R
Institut für Biochemie, Universität zu Köln, D-50674 Köln, Germany.
Morbach S
Krämer R
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