Lactococcus lactis uses MscL as its principal mechanosensitive channel

Joost H A Folgering, Paul C. Moe, Gea K. Schuurman-Wolters, Paul Blount, Bert Poolman

Research output: Contribution to journalArticle

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Abstract

The functions of the mechanosensitive channels from Lactococcus lactis were determined by biochemical, physiological, and electrophysiological methods. Patchclamp studies showed that the genes yncB and mscL encode MscS and MscL-like channels, respectively, when expressed in Escherichia coli or if the gene products were purified and reconstituted in proteoliposomes. However, unless yncB was expressed in trans, wild type membranes of L. lactis displayed only MscL activity. Membranes prepared from an mscL disruption mutant did not show any mechanosensitive channel activity, irrespective of whether the cells had been grown on low or high osmolarity medium. In osmotic downshift assays, wild type cells survived and retained 20% of the glycine betaine internalized under external high salt conditions. On the other hand, the mscL disruption mutant retained 40% of internalized glycine betaine and was significantly compromised in its survival upon osmotic downshifts. The data strongly suggest that L. lactis uses MscL as the main mechanosensitive solute release system to protect the cells under conditions of osmotic downshift.

Original languageEnglish (US)
Pages (from-to)8784-8792
Number of pages9
JournalJournal of Biological Chemistry
Volume280
Issue number10
DOIs
StatePublished - Mar 11 2005

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Betaine
Lactococcus lactis
Genes
Membranes
Escherichia coli
Assays
Salts
Osmolar Concentration
proteoliposomes

ASJC Scopus subject areas

  • Biochemistry

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Folgering, J. H. A., Moe, P. C., Schuurman-Wolters, G. K., Blount, P., & Poolman, B. (2005). Lactococcus lactis uses MscL as its principal mechanosensitive channel. Journal of Biological Chemistry, 280(10), 8784-8792. https://doi.org/10.1074/jbc.M411732200

Lactococcus lactis uses MscL as its principal mechanosensitive channel. / Folgering, Joost H A; Moe, Paul C.; Schuurman-Wolters, Gea K.; Blount, Paul; Poolman, Bert.

In: Journal of Biological Chemistry, Vol. 280, No. 10, 11.03.2005, p. 8784-8792.

Research output: Contribution to journalArticle

Folgering, JHA, Moe, PC, Schuurman-Wolters, GK, Blount, P & Poolman, B 2005, 'Lactococcus lactis uses MscL as its principal mechanosensitive channel', Journal of Biological Chemistry, vol. 280, no. 10, pp. 8784-8792. https://doi.org/10.1074/jbc.M411732200
Folgering, Joost H A ; Moe, Paul C. ; Schuurman-Wolters, Gea K. ; Blount, Paul ; Poolman, Bert. / Lactococcus lactis uses MscL as its principal mechanosensitive channel. In: Journal of Biological Chemistry. 2005 ; Vol. 280, No. 10. pp. 8784-8792.
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AB - The functions of the mechanosensitive channels from Lactococcus lactis were determined by biochemical, physiological, and electrophysiological methods. Patchclamp studies showed that the genes yncB and mscL encode MscS and MscL-like channels, respectively, when expressed in Escherichia coli or if the gene products were purified and reconstituted in proteoliposomes. However, unless yncB was expressed in trans, wild type membranes of L. lactis displayed only MscL activity. Membranes prepared from an mscL disruption mutant did not show any mechanosensitive channel activity, irrespective of whether the cells had been grown on low or high osmolarity medium. In osmotic downshift assays, wild type cells survived and retained 20% of the glycine betaine internalized under external high salt conditions. On the other hand, the mscL disruption mutant retained 40% of internalized glycine betaine and was significantly compromised in its survival upon osmotic downshifts. The data strongly suggest that L. lactis uses MscL as the main mechanosensitive solute release system to protect the cells under conditions of osmotic downshift.

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