Visualizing the self-assembly of tubulin with luminescent nanorods

被引:41
作者
Riegler, J
Nick, P
Kielmann, U
Nann, T [1 ]
机构
[1] Freiburg Mat Res Ctr, D-79104 Freiburg, Germany
[2] Univ Freiburg, Inst Biol 2, D-79104 Freiburg, Germany
关键词
fluorescent probes; semiconductor nanoparticles; self-assembly of tubulin; analytical methods; fluorescence microscopy; luminescence; colloids;
D O I
10.1166/jnn.2003.219
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
We succeeded in labeling tubulin with luminescent CdSe nanorods. The labeled tubulin was still able to self-assemble into microtubules, demonstrating that the protein has remained functional. The comparison with rhodamine-labeled tubulin revealed that whereas the rhodamine bleached completely within the observed time frame, the nanorods did not show any bleaching, which makes it possible to follow the key events of self-assembly at high time resolution. Preparation and derivatization of nanorods are described. Water-soluble, protein-reactive group-containing particles were covalently coupled to tubulin. The conjugate was purified by one cycle of assembly and disassembly and used to induce the formation of fluorescent microtubules. These results suggest that the use of luminescent nanorods should allow continuous confocal monitoring of dynamic biological processes.
引用
收藏
页码:380 / 385
页数:6
相关论文
共 20 条
[1]   Nanocrystal targeting in vivo [J].
Åkerman, ME ;
Chan, WCW ;
Laakkonen, P ;
Bhatia, SN ;
Ruoslahti, E .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2002, 99 (20) :12617-12621
[2]   Semiconductor nanocrystals as fluorescent biological labels [J].
Bruchez, M ;
Moronne, M ;
Gin, P ;
Weiss, S ;
Alivisatos, AP .
SCIENCE, 1998, 281 (5385) :2013-2016
[3]   Quantum dot bioconjugates for ultrasensitive nonisotopic detection [J].
Chan, WCW ;
Nie, SM .
SCIENCE, 1998, 281 (5385) :2016-2018
[4]   Trichosanthin induced calcium-dependent generation of reactive oxygen species in human choriocarcinoma cells [J].
Chun-Yang, Z ;
Yi-Xuan, G ;
Hui, M ;
Cheng-Cai, A ;
Die-Yan, C .
ANALYST, 2000, 125 (09) :1539-1542
[5]   Sorting fluorescent nanocrystals with DNA [J].
Gerion, D ;
Parak, WJ ;
Williams, SC ;
Zanchet, D ;
Micheel, CM ;
Alivisatos, AP .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2002, 124 (24) :7070-7074
[6]   Avidin: A natural bridge for quantum dot-antibody conjugates [J].
Goldman, ER ;
Balighian, ED ;
Mattoussi, H ;
Kuno, MK ;
Mauro, JM ;
Tran, PT ;
Anderson, GP .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2002, 124 (22) :6378-6382
[7]   Gravity-induced reorientation of cortical microtubules observed in vivo [J].
Himmelspach, R ;
Wymer, CL ;
Lloyd, CW ;
Nick, P .
PLANT JOURNAL, 1999, 18 (04) :449-453
[8]   Albumin-CdTe nanoparticle bioconjugates: Preparation, structure, and interunit energy transfer with antenna effect [J].
Mamedova, NN ;
Kotov, NA ;
Rogach, AL ;
Studer, J .
NANO LETTERS, 2001, 1 (06) :281-286
[9]   Self-assembly of CdSe-ZnS quantum dot bioconjugates using an engineered recombinant protein [J].
Mattoussi, H ;
Mauro, JM ;
Goldman, ER ;
Anderson, GP ;
Sundar, VC ;
Mikulec, FV ;
Bawendi, MG .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2000, 122 (49) :12142-12150
[10]   Programmed assembly of DNA functionalized quantum dots [J].
Mitchell, GP ;
Mirkin, CA ;
Letsinger, RL .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1999, 121 (35) :8122-8123