“Force-From-Lipids” Dependence of the MscCG Mechanosensitive Channel Gating on Anionic Membranes
Mechanosensory transduction in <i>Corynebacterium glutamicum</i> plays a major role in glutamate efflux for industrial MSG, whose production depends on the activation of MscCG-type mechanosensitive channels. Dependence of the MscCG channel activation by membrane tension on the membrane l...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-01-01
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| Series: | Microorganisms |
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| Online Access: | https://www.mdpi.com/2076-2607/11/1/194 |
| _version_ | 1797438428284977152 |
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| author | Yoshitaka Nakayama Paul R. Rohde Boris Martinac |
| author_facet | Yoshitaka Nakayama Paul R. Rohde Boris Martinac |
| author_sort | Yoshitaka Nakayama |
| collection | DOAJ |
| description | Mechanosensory transduction in <i>Corynebacterium glutamicum</i> plays a major role in glutamate efflux for industrial MSG, whose production depends on the activation of MscCG-type mechanosensitive channels. Dependence of the MscCG channel activation by membrane tension on the membrane lipid content has to date not been functionally characterized. Here, we report the MscCG channel patch clamp recording from liposomes fused with <i>C. glutamicum</i> membrane vesicles as well as from proteoliposomes containing the purified MscCG protein. Our recordings demonstrate that mechanosensitivity of MscCG channels depends significantly on the presence of negatively charged lipids in the proteoliposomes. MscCG channels in liposome preparations fused with native membrane vesicles exhibited the activation threshold similar to the channels recorded from <i>C. glutamicum</i> giant spheroplasts. In comparison, the activation threshold of the MscCG channels reconstituted into azolectin liposomes was higher than the activation threshold of <i>E. coli</i> MscL, which is gated by membrane tension close to the bilayer lytic tension. The spheroplast-like activation threshold was restored when the MscCG channels were reconstituted into liposomes made of <i>E. coli</i> polar lipid extract. In liposomes made of polar lipids mixed with synthetic phosphatidylethanolamine, phosphatidylglycerol, and cardiolipin, the activation threshold of MscCG was significantly reduced compared to the activation threshold recorded in azolectin liposomes, which suggests the importance of anionic lipids for the channel mechanosensitivity. Moreover, the micropipette aspiration technique combined with patch fluorometry demonstrated that membranes containing anionic phosphatidylglycerol are softer than membranes containing only polar non-anionic phosphatidylcholine and phosphatidylethanolamine. The difference in mechanosensitivity between <i>C. glutamicum</i> MscCG and canonical MscS of <i>E. coli</i> observed in proteoliposomes explains the evolutionary tuning of the force from lipids sensing in various bacterial membrane environments. |
| first_indexed | 2024-03-09T11:37:45Z |
| format | Article |
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| institution | Directory Open Access Journal |
| issn | 2076-2607 |
| language | English |
| last_indexed | 2024-03-09T11:37:45Z |
| publishDate | 2023-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Microorganisms |
| spelling | doaj.art-0ab661c7f03c4d4ca6068264d1b28a4c2023-11-30T23:37:47ZengMDPI AGMicroorganisms2076-26072023-01-0111119410.3390/microorganisms11010194“Force-From-Lipids” Dependence of the MscCG Mechanosensitive Channel Gating on Anionic MembranesYoshitaka Nakayama0Paul R. Rohde1Boris Martinac2Molecular Cardiology and Biophysics Division, Victor Chang Cardiac Research Institute, Sydney 2010, AustraliaMolecular Cardiology and Biophysics Division, Victor Chang Cardiac Research Institute, Sydney 2010, AustraliaMolecular Cardiology and Biophysics Division, Victor Chang Cardiac Research Institute, Sydney 2010, AustraliaMechanosensory transduction in <i>Corynebacterium glutamicum</i> plays a major role in glutamate efflux for industrial MSG, whose production depends on the activation of MscCG-type mechanosensitive channels. Dependence of the MscCG channel activation by membrane tension on the membrane lipid content has to date not been functionally characterized. Here, we report the MscCG channel patch clamp recording from liposomes fused with <i>C. glutamicum</i> membrane vesicles as well as from proteoliposomes containing the purified MscCG protein. Our recordings demonstrate that mechanosensitivity of MscCG channels depends significantly on the presence of negatively charged lipids in the proteoliposomes. MscCG channels in liposome preparations fused with native membrane vesicles exhibited the activation threshold similar to the channels recorded from <i>C. glutamicum</i> giant spheroplasts. In comparison, the activation threshold of the MscCG channels reconstituted into azolectin liposomes was higher than the activation threshold of <i>E. coli</i> MscL, which is gated by membrane tension close to the bilayer lytic tension. The spheroplast-like activation threshold was restored when the MscCG channels were reconstituted into liposomes made of <i>E. coli</i> polar lipid extract. In liposomes made of polar lipids mixed with synthetic phosphatidylethanolamine, phosphatidylglycerol, and cardiolipin, the activation threshold of MscCG was significantly reduced compared to the activation threshold recorded in azolectin liposomes, which suggests the importance of anionic lipids for the channel mechanosensitivity. Moreover, the micropipette aspiration technique combined with patch fluorometry demonstrated that membranes containing anionic phosphatidylglycerol are softer than membranes containing only polar non-anionic phosphatidylcholine and phosphatidylethanolamine. The difference in mechanosensitivity between <i>C. glutamicum</i> MscCG and canonical MscS of <i>E. coli</i> observed in proteoliposomes explains the evolutionary tuning of the force from lipids sensing in various bacterial membrane environments.https://www.mdpi.com/2076-2607/11/1/194bacterial mechanosensingbacterial electrophysiologypatch fluorometrymicropipette aspirationmembrane stiffness<i>Corynebacterium glutamicum</i> |
| spellingShingle | Yoshitaka Nakayama Paul R. Rohde Boris Martinac “Force-From-Lipids” Dependence of the MscCG Mechanosensitive Channel Gating on Anionic Membranes Microorganisms bacterial mechanosensing bacterial electrophysiology patch fluorometry micropipette aspiration membrane stiffness <i>Corynebacterium glutamicum</i> |
| title | “Force-From-Lipids” Dependence of the MscCG Mechanosensitive Channel Gating on Anionic Membranes |
| title_full | “Force-From-Lipids” Dependence of the MscCG Mechanosensitive Channel Gating on Anionic Membranes |
| title_fullStr | “Force-From-Lipids” Dependence of the MscCG Mechanosensitive Channel Gating on Anionic Membranes |
| title_full_unstemmed | “Force-From-Lipids” Dependence of the MscCG Mechanosensitive Channel Gating on Anionic Membranes |
| title_short | “Force-From-Lipids” Dependence of the MscCG Mechanosensitive Channel Gating on Anionic Membranes |
| title_sort | force from lipids dependence of the msccg mechanosensitive channel gating on anionic membranes |
| topic | bacterial mechanosensing bacterial electrophysiology patch fluorometry micropipette aspiration membrane stiffness <i>Corynebacterium glutamicum</i> |
| url | https://www.mdpi.com/2076-2607/11/1/194 |
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