DOI: 10.1126/science.1091749. Biomolecular Interactions at Phospholipid-Decorated Surfaces of Liquid Crystals

Biomolecular Interactions at Phospholipid-Decorated Surfaces of Liquid Crystals

10.1126/science.1091749

Crossref journal-article

American Association for the Advancement of Science (AAAS)

Science (221)

Abstract

The spontaneous assembly of phospholipids at planar interfaces between thermotropic liquid crystals and aqueous phases gives rise to patterned orientations of the liquid crystals that reflect the spatial and temporal organization of the phospholipids. Strong and weak specific-binding events involving proteins at these interfaces drive the reorganization of the phospholipids and trigger orientational transitions in the liquid crystals. Because these interfaces are fluid, processes involving the lateral organization of proteins (such as the formation of protein- and phospholipid-rich domains) are also readily imaged by the orientational response of the liquid crystal, as are stereospecific enzymatic events. These results provide principles for label-free monitoring of aqueous streams for molecular and biomolecular species without the need for complex instrumentation.

Bibliography

Brake, J. M., Daschner, M. K., Luk, Y.-Y., & Abbott, N. L. (2003). Biomolecular Interactions at Phospholipid-Decorated Surfaces of Liquid Crystals. Science, 302(5653), 2094â2097.

Authors 4

Jeffrey M. Brake (first)
Maren K. Daschner (additional)
Yan-Yeung Luk (additional)
Nicholas L. Abbott (additional)

References 34 Referenced 582

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R. V. Stahelin, W. Cho, Biochem. J.359, 679 (2001). (10.1042/bj3590679) / Biochem. J. (2001)
C. Dietrich et al., Biophys. J.80, 1417 (2001). (10.1016/S0006-3495(01)76114-0) / Biophys. J. (2001)
A. P. Gast, C. R. Robertson, S.-W. Wang, M. T. Yatcilla, Biomol. Eng.16, 21 (1999). (10.1016/S1050-3862(99)00045-5) / Biomol. Eng. (1999)
10.1126/science.8342021
R. A. Walker, J. C. Conboy, G. L. Richmond, Langmuir13, 3070 (1997). (10.1021/la961007q) / Langmuir (1997)
E. Sackmann, Science271, 43 (1996). (10.1126/science.271.5245.43) / Science (1996)
10.1126/science.275.5300.651
N. M. Rao, A. L. Plant, V. Silin, S. Wight, S. W. Hui, Biophys. J.73, 3066 (1997). (10.1016/S0006-3495(97)78334-6) / Biophys. J. (1997)
T. Yang, S.-Y. Jung, H. Mao, P. S. Cremer, Anal. Chem.73, 165 (2001). (10.1021/ac000997o) / Anal. Chem. (2001)
K. Tanaka, P. A. Manning, H. Yu, Langmuir16, 2665 (2000). (10.1021/la9909244) / Langmuir (2000)
Previous studies have reported the use of LCs to image proteins bound to solid surfaces ( 13 ).
V. K. Gupta, J. J. Skaife, T. B. Dubrovsky, N. L. Abbott, Science279, 2077 (1998). (10.1126/science.279.5359.2077) / Science (1998)
Materials and methods are available as supporting material on Science Online.
The temporal characteristics (∼0.5 to 2 hours) of the process of transfer of L-DLPC revealed by the LC are similarto previous studies of the fusion of vesicles with hydrophobic surfaces ( 17 32 ).
K. Hiltrop, H. Stegemeyer, Ber. Bunsenges. Phys. Chem.85, 582 (1981). (10.1002/bbpc.19810850712) / Ber. Bunsenges. Phys. Chem. (1981)
E. Kalb, S. Frey, L. K. Tamm, Biochim. Biophys. Acta1103, 307 (1992). (10.1016/0005-2736(92)90101-Q) / Biochim. Biophys. Acta (1992)
Whereas L-DLPC within vesicles is fluid and mobile at room temperature and thus readily transfers to form monolayers at hydrophobic interfaces D-DPPC does not. By using fluorescence microscopy we confirmed that the areal density of D-DPPC transferred to the aqueous-LC interface from vesicles was only ∼70% of that observed for L-DLPC. However by using mixed micelles formed from DTAB and D-DPPC we demonstrated that it was possible to prepare LC interfaces laden with D-DPPC with areal densities similarto the L-DLPC interfaces prepared from vesicles ( 33 ). D-DPPC transferred under these conditions also caused a perpendicular orientation of the LC at the aqueous-LC interface thus establishing a general procedure for assembling lipids at the surface of LCs.
U. Dahmen-Levison, G. Brezesinski, H. Muhwald, Thin Solid Films616, 327 (1998). / Thin Solid Films (1998)
The lateral diffusivity of TR-DHPE within the lipid layer was measured with quantitative fluorescence imaging (photobleaching) to be 6 × 10 –8 cm 2 /s ( 21 ) consistent with measurements of the diffusivities of lipids at a variety of interfaces including biological membranes ( 17 ). Fluorescence imaging also revealed compartmentalization of the lipid between grid squares which we have exploited to form patterned arrays of lipids at these interfaces (stability > 1 week).
The lipid diffusivity was measured in photobleaching experiments using a confocal microscope (Keck Bioimaging Facility at University of Wisconsin). A step-edge bleaching profile was formed and the recovery was imaged over time. At short times the recovery profile c ( x t ) could be modeled as a 1D diffusion problem following the equation c ( x t ) = erf [ x /(4 Dt ) 0.5 ] where x is position t is time erf is the error function and the diffusivity D was the only fitting parameter.
PLA 2 is found in the venom of a variety of animals (including snakes) and facilitates the passage of toxins across cell membranes.
P. P. M. Bonsen, G. H. de Haas, W. A. Pieterson, L. L. M. van Deenen, Biochim. Biophys. Acta270, 364 (1972). (10.1016/0005-2760(72)90200-7) / Biochim. Biophys. Acta (1972)
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G. H. de Haas, P. P. M. Bonsen, W. A Pieterson, L. L. M. van Deenen, Biochim. Biophys. Acta239, 252 (1971). (10.1016/0005-2760(71)90171-8) / Biochim. Biophys. Acta (1971)
G. Zografi, R. Verger, G. H. de Haas, Chem. Phys. Lipids7, 185 (1971). (10.1016/0009-3084(71)90001-6) / Chem. Phys. Lipids (1971)
Reaction time-scale calculations assume a steady-state hydrolysis rate expressed as v = Q m E 0 Γ S where v is the reaction velocity (in molecules cm –2 s –1 ) Q m is the global kinetic constant of hydrolysis of monolayers of DLPC by PLA 2 (∼10 –15 cm 3 permolecule persecond) E 0 is the total enzyme concentration (∼6 × 10 11 molecules/cm 3 ) and Γ S is the surface concentration of DLPC ( 30 ). The reaction time t can be estimated from the integrated form of this equation t = ( Q m E 0 ) –1 ln (1/ X ) where X is the fraction of remaining substrate. Conversions from 63 to 95% of DLPC into products ( X = 0.37 to 0.05) correspond to the time scales observed in the orientational response of the LC. The diffusion time scale forPLA 2 to the interface is estimated to be ∼11 s [ t ∼ l 2 / D where l is the diffusion length (∼33 μm) and D ∼ 10 –6 cm 2 /s].
S. Ransac, M. Ivanova, R. Verger, I. Panaiotov, Methods Enyzmol.286, 263 (1997). / Methods Enyzmol. (1997)
D. W. Grainger et al., J. Controlled Release19, 201 (1992). (10.1016/0168-3659(92)90077-5) / J. Controlled Release (1992)
J. B. Hubbard, V. Silin, A. L. Plant, Biophys. Chem.75, 163 (1998). (10.1016/S0301-4622(98)00199-9) / Biophys. Chem. (1998)
L. M. Grant, F. Tiberg, Biophys. J.82, 1373 (2002). (10.1016/S0006-3495(02)75492-1) / Biophys. J. (2002)
Supported by NSF NIH and the U.S. Office of Naval Research.

Dates

Type	When
Created	21 years, 8 months ago (Dec. 18, 2003, 5:57 p.m.)
Deposited	1 year, 7 months ago (Jan. 9, 2024, 9:12 p.m.)
Indexed	51 minutes ago (Aug. 27, 2025, 2:37 p.m.)
Issued	21 years, 8 months ago (Dec. 19, 2003)
Published	21 years, 8 months ago (Dec. 19, 2003)
Published Print	21 years, 8 months ago (Dec. 19, 2003)

Funders 0

None

BibTeX

@article{Brake_2003, title={Biomolecular Interactions at Phospholipid-Decorated Surfaces of Liquid Crystals}, volume={302}, ISSN={1095-9203}, url={http://dx.doi.org/10.1126/science.1091749}, DOI={10.1126/science.1091749}, number={5653}, journal={Science}, publisher={American Association for the Advancement of Science (AAAS)}, author={Brake, Jeffrey M. and Daschner, Maren K. and Luk, Yan-Yeung and Abbott, Nicholas L.}, year={2003}, month=dec, pages={2094–2097} }

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