In vitro sliding of actin filaments labelled with single quantum dots

被引:102
作者
Månsson, A
Sundberg, M
Balaz, M
Bunk, R
Nicholls, IA
Omling, P
Tågerud, S
Montelius, L
机构
[1] Univ Kalmar, Dept Chem & Biomed Sci, SE-39182 Kalmar, Sweden
[2] Lund Univ, Div Solid State Phys, SE-22100 Lund, Sweden
[3] Lund Univ, Nanometre Consortium, SE-22100 Lund, Sweden
关键词
myosin; actin; quantum dot; nanostructure; nanocrystal; motility assay;
D O I
10.1016/j.bbrc.2003.12.133
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We recently refined the in vitro motility assay for studies of actomyosin function to achieve rectified myosin induced sliding of actin filaments. This paves the way, both for detailed functional studies of actomyosin and for nanotechnological applications. In the latter applications it would be desirable to use actin filaments for transportation of cargoes (e.g., enzymes) between different predetermined locations on a chip. We here describe how single quantum dot labelling of isolated actin filaments simultaneously provides handles for cargo attachment and bright and photostable fluorescence labels facilitating cargo detection and filament tracking. Labelling was achieved with preserved actomyosin function using streptavidin-coated CdSe quantum dots (Qdots). These nanocrystals have several unique physical properties and the present work describes their first use for functional studies of isolated proteins outside the cell. The results, in addition to the nanotechnology developments, open for new types of in vitro assays of isolated biomolecules. (C) 2003 Elsevier Inc. All rights reserved.
引用
收藏
页码:529 / 534
页数:6
相关论文
共 32 条
[1]   Natural strategies for the molecular engineer [J].
Ball, P .
NANOTECHNOLOGY, 2002, 13 (05) :R15-R28
[2]   ADP binding induces an asymmetry between the heads of unphosphorylated myosin [J].
Berger, CEM ;
Fagnant, PM ;
Heizmann, S ;
Trybus, KM ;
Geeves, MA .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2001, 276 (26) :23240-23245
[3]   Quantum dot research: Current state and future prospects [J].
Bukowski, TJ ;
Simmons, JH .
CRITICAL REVIEWS IN SOLID STATE AND MATERIALS SCIENCES, 2002, 27 (3-4) :119-142
[4]   Towards a 'nano-traffic' system powered by molecular motors [J].
Bunk, R ;
Klinth, J ;
Rosengren, J ;
Nicholls, I ;
Tågerud, S ;
Omling, P ;
Månsson, A ;
Montelius, L .
MICROELECTRONIC ENGINEERING, 2003, 67-8 :899-904
[5]   Actomyosin motility on nanostructured surfaces [J].
Bunk, R ;
Klinth, J ;
Montelius, L ;
Nicholls, IA ;
Omling, P ;
Tågerud, S ;
Månsson, A .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2003, 301 (03) :783-788
[6]   Quantum dot bioconjugates for ultrasensitive nonisotopic detection [J].
Chan, WCW ;
Nie, SM .
SCIENCE, 1998, 281 (5385) :2016-2018
[7]   Diffusion dynamics of glycine receptors revealed by single-quantum dot tracking [J].
Dahan, M ;
Lévi, S ;
Luccardini, C ;
Rostaing, P ;
Riveau, B ;
Triller, A .
SCIENCE, 2003, 302 (5644) :442-445
[8]  
Empedocles SA, 1999, ADV MATER, V11, P1243, DOI 10.1002/(SICI)1521-4095(199910)11:15<1243::AID-ADMA1243>3.0.CO
[9]  
2-2
[10]   Photoluminescence spectroscopy of single CdSe nanocrystallite quantum dots [J].
Empedocles, SA ;
Norris, DJ ;
Bawendi, MG .
PHYSICAL REVIEW LETTERS, 1996, 77 (18) :3873-3876