Hostname: page-component-586b7cd67f-dlnhk Total loading time: 0 Render date: 2024-11-24T12:30:04.738Z Has data issue: false hasContentIssue false
  • English
  • Français

The Gating Mechanism of Mechanosensitive Channels in Droplet Interface Bilayers

Published online by Cambridge University Press:  16 June 2015

Joseph S. Najem ,
Eric Freeman ,
Sergei Sukharev  and
Donald J. Leo
Joseph S. Najem
Affiliation:
Department of Mechanical Engineering, Virginia Polytechnic Institute and State University, Blacksburg, VA 24061, U.S.A.
Eric Freeman
Affiliation:
College of Engineering, University of Georgia, Athens, GA 30602, U.S.A.
Sergei Sukharev
Affiliation:
Department of Biology, University of Maryland, College Park, MD 20742, U.S.A
Donald J. Leo
Affiliation:
College of Engineering, University of Georgia, Athens, GA 30602, U.S.A.
Get access

Abstract

MscL, a large-conductance mechanosensitive channel, is a ubiquitous osmolyte release valve that aids bacteria in surviving abrupt hypo-osmotic shocks. The large scale of its tension-driven opening transition makes it a strong candidate to serve as a transducer in novel stimuli-responsive biomolecular materials. In the previous work, a low-threshold gain-of-function V23T mutant of MscL produced a reliable activation behavior in a droplet interface bilayer (DIB) with applied axial droplet compression. Near the maximal compression, the aqueous droplets deform and the resulting increase in surface area leads to an increase in tension in the water-lipid-oil interface. This increase in tension is the product of the relative change in the droplet surface area and the elastic modulus of the DPhPC lipid monolayer (∼120 mN/m). This paper, presents a study of the physical processes that cause MscL gating in the DIB. Analysis of video during compression and relaxation of the droplets is utilized to estimate the change in the surface area of the droplet and the variation on monolayer surface tension. The monolayer surface tension is proportional to the area change of the droplet normalized to the original surface area. The results demonstrate that the area change in the droplet is negligible at frequencies above 1 Hz, but is approximately 2% at frequencies in the range of 100 mHz. In addition, at low frequencies (∼0.2 Hz) bilayer thinning occurs at maximum compression, proving an increase in bilayer tension. However, this study also shows that gating at frequencies higher than 0.2 Hz could be achieved through the application of high duty cycle oscillation (∼75%). The relative change in monolayer area increases significantly at higher duty cycle oscillations where the compression stroke is much faster than the relaxation stroke.

Keywords

biologicalbiological synthesis (assembly)self-assembly
Type
Articles
Information
MRS Online Proceedings Library (OPL) , Volume 1722: Symposium F – Reverse Engineering of Bioinspired Nanomaterials , 2015 , mrsf14-1722-f05-49
Copyright
Copyright © Materials Research Society 2015 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Perozo, E., NAT REV MOL CELL BIO 7 (2), 109–119 (2006).CrossRefGoogle Scholar
Kung, C., Martinac, B. and Sukharev, S., ANNU REV MICROBIOL 64, 313–329 (2010).CrossRefGoogle Scholar
Sukharev, S. I., Martinac, B., Arshavsky, V. Y. and Kung, C., Biophysical Journal 65 (1), 177–183 (1993).CrossRefGoogle Scholar
Najem, J., Dunlap, M., Sukharev, S. and Leo, D. J., MRS Proceedings 1621, mrsf13-1621-d1601-1605 (2014).CrossRefGoogle Scholar
Tien, H. T., Bilayer lipid membranes (BLM): theory and practice. (M. Dekker, 1974).Google Scholar
Andersson, M., Okeyo, G., Wilson, D., Keizer, H., Moe, P., Blount, P., Fine, D., Dodabalapur, A. and Duran, R. S., Biosensors and Bioelectronics 23 (6), 919–923 (2008).CrossRefGoogle Scholar
Barriga, H. M., Booth, P., Haylock, S., Bazin, R., Templer, R. H. and Ces, O., J R Soc Interface 11 (98), 20140404 (2014).CrossRefGoogle Scholar
Sukharev, S., Durell, S. R. and Guy, H. R., BIOPHYS J 81 (2), 917–936 (2001).CrossRefGoogle Scholar
Sarles, S. A. and Leo, D. J., ANAL CHEM 82 (3), 959–966 (2010).CrossRefGoogle Scholar
Anishkin, A., Chiang, C.-S. and Sukharev, S., J GEN PHYSIOL 125 (2), 155–170 (2005).CrossRefGoogle Scholar
Li, Y., Moe, P. C., Chandrasekaran, S., Booth, I. R. and Blount, P., The EMBO journal 21 (20), 5323–5330 (2002).CrossRefGoogle Scholar
Levina, N., Tötemeyer, S., Stokes, N. R., Louis, P., Jones, M. A. and Booth, I. R., The EMBO Journal 18 (7), 1730–1737 (1999).CrossRefGoogle Scholar
Sukharev, S. I., Sigurdson, W. J., Kung, C. and Sachs, F., J GEN PHYSIOL 113 (4), 525–540 (1999).CrossRefGoogle Scholar
Chiang, C.-S., Anishkin, A. and Sukharev, S., BIOPHYS J 86 (5), 2846–2861 (2004).CrossRefGoogle Scholar
Yasmann, A. and Sukharev, S., Langmuir, submitted (2014).Google Scholar