ABSTRACT The peptide GsMTx4, isolated from the venom of the tarantula _Grammostola spatulata_, is a selective inhibitor of stretch-activated cation channels (SACs)1. The mechanism of

inhibition remains unknown; but both GsMTx4 and its enantiomer, enGsMTx4, modify the gating of SACs, thus violating a trademark of the traditional lock-and-key model of ligand–protein

interactions. Suspecting a bilayer-dependent mechanism, we examined the effect of GsMTx4 and enGsMTx4 on gramicidin A (gA) channel gating2. Both peptides are active, and the effect increases

with the degree of hydrophobic mismatch between bilayer thickness and channel length, meaning that GsMTx4 decreases the energy required to deform the boundary lipids adjacent to the

peptides—mechanopharmacology—involves not only the protein itself but also the surrounding lipids. The surprising efficacy of the d form of GsMTx4 peptide has important therapeutic

implications, because d peptides are not hydrolysed by endogenous proteases and may be administered orally. Access through your institution Buy or subscribe This is a preview of subscription

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OF SMALL-MOLECULE MODULATION OF THE GATING OF ACID-SENSING ION CHANNEL 1 Article Open access 09 February 2021 STRUCTURAL BASIS FOR EXCITATORY NEUROPEPTIDE SIGNALING Article Open access 09

February 2024 STRUCTURAL BASIS OF INHIBITION OF HUMAN NAV1.8 BY THE TARANTULA VENOM PEPTIDE PROTOXIN-I Article Open access 07 February 2025 REFERENCES * Suchyna, T. M. et al. Identification

of a peptide toxin from _Grammostola spatulata_ spider venom that blocks cation-selective stretch-activated channels. _J. Gen. Physiol._ 115, 583–598 (2000) Article  CAS  PubMed  PubMed

Central  Google Scholar  * Andersen, O. S. et al. Ion channels as tools to monitor lipid bilayer-membrane protein interactions: gramicidin channels as molecular force transducers. _Methods

Enzymol._ 294, 208–224 (1999) Article  CAS  PubMed  Google Scholar  * Patel, A. J. et al. Inhalational anesthetics activate two-pore-domain background K+ channels. _Nature Neurosci._ 2,

422–426 (1999) Article  CAS  PubMed  Google Scholar  * Patel, A. J., Lazdunski, M. & Honore, E. Lipid and mechano-gated 2P domain K+ channels. _Curr. Opin. Cell Biol._ 13, 422–427 (2001)

Article  CAS  PubMed  Google Scholar  * Perozo, E., Kloda, A., Cortes, D. M. & Martinac, B. Physical principles underlying the transduction of bilayer deformation forces during

mechanosensitive channel gating. _Nature Struct. Biol._ 9, 696–703 (2002) Article  CAS  PubMed  Google Scholar  * Lundbæk, J. A. & Andersen, O. S. Lysophospholipids modulate channel

function by altering the mechanical properties of lipid bilayers. _J. Gen. Physiol._ 104, 645–673 (1994) Article  PubMed  Google Scholar  * Hwang, T. C., Koeppe, R. E. II & Andersen, O.

S. Genistein can modulate channel function by a phosphorylation-independent mechanism: importance of hydrophobic mismatch and bilayer mechanics. _Biochemistry_ 42, 13646–13658 (2003) Article

  CAS  PubMed  Google Scholar  * Goulian, M. et al. Gramicidin channel kinetics under tension. _Biophys. J._ 74, 328–337 (1998) Article  ADS  CAS  PubMed  PubMed Central  Google Scholar  *

Oswald, R. E., Suchyna, T. M., McFeeters, R., Gottlieb, P. & Sachs, F. Solution structure of peptide toxins that block mechanosensitive ion channels. _J. Biol. Chem._ 277, 34443–34450

(2002) Article  CAS  PubMed  Google Scholar  * Ostrow, K. L. et al. cDNA sequence and _in vitro_ folding of GsMTx4, a specific peptide inhibitor of mechanosensitive channels. _Toxicon_ 42,

263–274 (2003) Article  CAS  PubMed  Google Scholar  * Markin, V. S. & Sachs, F. Thermodynamics of mechanosensitivity. _Physical Biol._ (in the press) * Suchyna, T. & Sachs, F.

Dynamic regulation of mechanosensitive channels: capacitance used to monitor patch tension in real time. _Phys. Biol._ 1, 1–18 (2004) Article  ADS  CAS  PubMed  Google Scholar  * Ladokhin,

A. S., Jayasinghe, S. & White, S. H. How to measure and analyze tryptophan fluorescence in membranes properly, and why bother? _Anal. Biochem._ 285, 235–245 (2000) Article  CAS  PubMed 

Google Scholar  * White, S. H., Wimley, W. C., Ladokhin, A. S. & Hristova, K. Protein folding in membranes: determining energetics of peptide-bilayer interactions. _Methods Enzymol._

295, 62–87 (1998) Article  CAS  PubMed  Google Scholar  * Kim, J., Mosior, M., Chung, L. A., Wu, H. & McLaughlin, S. Binding of peptides with basic residues to membranes containing

acidic phospholipids. _Biophys. J._ 60, 135–148 (1991) Article  CAS  PubMed  PubMed Central  Google Scholar  * Lundbæk, J. A. & Andersen, O. S. Spring constants for channel-induced lipid

bilayer deformations—estimates using gramicidin channels. _Biophys. J._ 76, 889–895 (1999) Article  PubMed  PubMed Central  Google Scholar  * O'Connell, A. M., Koeppe, R. E. II &

Andersen, O. S. Kinetics of gramicidin channel formation in lipid bilayers: transmembrane monomer association. _Science_ 250, 1256–1259 (1990) Article  ADS  CAS  PubMed  Google Scholar  *

Elliott, J. R., Needham, D., Dilger, J. P. & Haydon, D. A. The effects of bilayer thickness and tension on gramicidin single-channel lifetime. _Biochim. Biophys. Acta_ 735, 95–103 (1983)

Article  CAS  PubMed  Google Scholar  * Huang, H. W. Deformation free energy of bilayer membrane and its effect on gramicidin channel lifetime. _Biophys. J._ 50, 1061–1070 (1986) Article 

ADS  CAS  PubMed  PubMed Central  Google Scholar  * Koeppe, R. E. II et al. On the helix sense of gramicidin A single channels. _Proteins_ 12, 49–62 (1992) Article  CAS  PubMed  Google

Scholar  * Trudelle, Y. & Heitz, F. Synthesis and characterization of Tyr(Bzl)9,11,13,15 and Tyr9,11,13,15 gramicidin A. _Int. J. Pept. Protein Res._ 30, 163–169 (1987) Article  CAS 

PubMed  Google Scholar  * Lundbæk, J. A. et al. Regulation of sodium channel function by bilayer elasticity the importance of hydrophobic coupling: effects of micelle-forming amphiphiles and

cholesterol. _J. Gen. Physiol._ 123, 599–621 (2004) Article  PubMed  PubMed Central  Google Scholar  * Bode, F., Sachs, F. & Franz, M. R. Tarantula peptide inhibits atrial fibrillation.

_Nature_ 409, 35–36 (2001) Article  ADS  CAS  PubMed  Google Scholar  * Lehtonen, J. Y. & Kinnunen, P. K. Phospholipase A2 as a mechanosensor. _Biophys. J._ 68, 1888–1894 (1995) Article

  ADS  CAS  PubMed  PubMed Central  Google Scholar  * Gudi, S., Nolan, J. P. & Frangos, J. A. Modulation of GTPase activity of G proteins by fluid shear stress and phospholipid

composition. _Proc. Natl Acad. Sci. USA_ 95, 2515–2519 (1998) Article  ADS  CAS  PubMed  PubMed Central  Google Scholar  * Laitko, U. & Morris, C. E. Membrane tension accelerates

rate-limiting voltage-dependent activation and slow inactivation steps in a _Shaker_ channel. _J. Gen. Physiol._ 123, 135–154 (2004) Article  PubMed  PubMed Central  Google Scholar  *

Greathouse, D. V., Koeppe, R. E. II, Providence, L. L., Shobana, S. & Andersen, O. S. Design and characterization of gramicidin channels. _Methods Enzymol._ 294, 525–550 (1999) Article 

CAS  PubMed  Google Scholar  * Andersen, O. S. Ion movement through gramicidin A channels. Single-channel measurements at very high potentials. _Biophys. J._ 41, 119–133 (1983) Article  ADS

  CAS  PubMed  PubMed Central  Google Scholar  * Bett, G. C. & Sachs, F. Activation and inactivation of mechanosensitive currents in the chick heart. _J. Membr. Biol._ 173, 237–254

(2000) Article  CAS  PubMed  Google Scholar  Download references ACKNOWLEDGEMENTS We would like to thank J. Niggel (SUNY) and A. Ladokhin (UC Irvine) for the fluorescence data, and M.

Teeling for the tissue culture. This work is supported by NIH grants to O.S.A. and F.S. AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * Department of Physiology and Biophysics, SUNY at

Buffalo, Buffalo, New York, 14214, USA Thomas M. Suchyna, Frederick Sachs & Philip A. Gottlieb * Department of Physiology and Biophysics, Weill Medical College of Cornell University, New

York, New York, 10021, USA Sonya E. Tape & Olaf S. Andersen * Department of Chemistry and Biochemistry, University of Arkansas, Fayetteville, Arkansas, 72701, USA Roger E. Koeppe II

Authors * Thomas M. Suchyna View author publications You can also search for this author inPubMed Google Scholar * Sonya E. Tape View author publications You can also search for this author

inPubMed Google Scholar * Roger E. Koeppe II View author publications You can also search for this author inPubMed Google Scholar * Olaf S. Andersen View author publications You can also

search for this author inPubMed Google Scholar * Frederick Sachs View author publications You can also search for this author inPubMed Google Scholar * Philip A. Gottlieb View author

publications You can also search for this author inPubMed Google Scholar CORRESPONDING AUTHOR Correspondence to Olaf S. Andersen. ETHICS DECLARATIONS COMPETING INTERESTS The authors declare

that they have no competing financial interests. RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Suchyna, T., Tape, S., Koeppe, R. _et al._

Bilayer-dependent inhibition of mechanosensitive channels by neuroactive peptide enantiomers. _Nature_ 430, 235–240 (2004). https://doi.org/10.1038/nature02743 Download citation * Received:

09 April 2004 * Accepted: 03 June 2004 * Published: 08 July 2004 * Issue Date: 08 July 2004 * DOI: https://doi.org/10.1038/nature02743 SHARE THIS ARTICLE Anyone you share the following link

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