Kinetics of DNA-mediated docking reactions between vesicles tethered to supported lipid bilayers

被引:72
作者
Chan, Yee-Hung M.
Lenz, Peter
Boxer, Steven G.
机构
[1] Stanford Univ, Dept Chem, Stanford, CA 94305 USA
[2] Univ Marburg, Dept Phys, D-35032 Marburg, Germany
关键词
D O I
10.1073/pnas.0706114104
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Membrane-membrane recognition and binding are crucial in many biological processes. We report an approach to studying the dynamics of such reactions by using DNA-tethered vesicles as a general scaffold for displaying membrane components. This system was used to characterize the docking reaction between two populations of tethered vesicles that display complementary DNA. Deposition of vesicles onto a supported lipid bilayer was performed by using a microfluidic device to prevent mixing of the vesicles in bulk during sample preparation. Once tethered onto the surface, vesicles mixed via two-dimensional diffusion. DNA-mediated docking of two reacting vesicles results in their colocalization after collision and their subsequent tandem motion. Individual docking events and population kinetics were observed via epifluorescence microscopy. A lattice-diffusion simulation was implemented to extract from experimental data the probability, P-dock, that a collision leads to docking. For individual vesicles displaying small numbers of docking DNA, Pdock shows a first-order relationship with copy number as well as a strong dependence on the DNA sequence. Both trends are explained by a model that includes both tethered vesicle diffusion on the supported bilayer and docking DNA diffusion over each vesicle's surface. These results provide the basis for the application of tethered vesicles to study other membrane reactions including protein-mediated docking and fusion.
引用
收藏
页码:18913 / 18918
页数:6
相关论文
共 36 条
[1]   Probing the structure of supported membranes and tethered oligonucleotides by fluorescence interference contrast microscopy [J].
Ajo-Franklin, CM ;
Yoshina-Ishii, C ;
Boxer, SG .
LANGMUIR, 2005, 21 (11) :4976-4983
[2]   Membrane on a chip:: A functional tethered lipid bilayer membrane on silicon oxide surfaces [J].
Atanasov, V ;
Knorr, N ;
Duran, RS ;
Ingebrandt, S ;
Offenhäusser, A ;
Knoll, W ;
Köper, I .
BIOPHYSICAL JOURNAL, 2005, 89 (03) :1780-1788
[3]   Immobilization in surface-tethered lipid vesicles as a new tool for single biomolecule spectroscopy [J].
Boukobza, E ;
Sonnenfeld, A ;
Haran, G .
JOURNAL OF PHYSICAL CHEMISTRY B, 2001, 105 (48) :12165-12170
[4]   Single molecule observation of liposome-bilayer fusion thermally induced by soluble N-ethyl maleimide sensitive-factor attachment protein receptors (SNAREs) [J].
Bowen, ME ;
Weninger, K ;
Brunger, AT ;
Chu, S .
BIOPHYSICAL JOURNAL, 2004, 87 (05) :3569-3584
[5]   ALLOGENEIC STIMULATION OF CYTO-TOXIC T-CELLS BY SUPPORTED PLANAR MEMBRANES [J].
BRIAN, AA ;
MCCONNELL, HM .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1984, 81 (19) :6159-6163
[6]   SNARE-mediated lipid mixing depends on the physical state of the vesicles [J].
Chen, XC ;
Araç, D ;
Wang, TM ;
Gilpin, CJ ;
Zimmerberg, J ;
Rizo, J .
BIOPHYSICAL JOURNAL, 2006, 90 (06) :2062-2074
[7]   Measuring two-dimensional receptor-ligand binding kinetics by micropipette [J].
Chesla, SE ;
Selvaraj, P ;
Zhu, C .
BIOPHYSICAL JOURNAL, 1998, 75 (03) :1553-1572
[8]   Neuronal SNAREs do not trigger fusion between synthetic membranes but do promote PEG-mediated membrane fusion [J].
Dennison, SM ;
Bowen, ME ;
Brunger, AT ;
Lentz, BR .
BIOPHYSICAL JOURNAL, 2006, 90 (05) :1661-1675
[9]   Membranes as messengers in T cell adhesion signaling [J].
Dustin, ML ;
Bivona, TG ;
Philips, MR .
NATURE IMMUNOLOGY, 2004, 5 (04) :363-372
[10]   Imaging single membrane fusion events mediated by SNARE proteins [J].
Fix, M ;
Melia, TJ ;
Jaiswal, JK ;
Rappoport, JZ ;
You, DQ ;
Söllner, TH ;
Rothman, JE ;
Simon, SM .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2004, 101 (19) :7311-7316