Abstract
Cytoplasmic dynein, which is the largest and arguably the most complex cytoskeletal motor protein, plays fundamental roles during cell division, nuclear positioning, and organelle and mRNA transport, by generating force and movement toward the minus ends of microtubules. Consequently, dynein is central to many physiological processes, and its dysfunction is implicated in human diseases. However, the molecular mechanism by which dynein produces force and movement remains poorly understood. Here, we describe the use of optical tweezers to probe the nanometer-scale motion and force generation of individual dynein molecules, and provide a hands-on protocol for how to purify cytoplasmic dynein from budding yeast in amounts sufficient for single-molecule studies.
| Original language | English (US) |
|---|---|
| Title of host publication | Methods in Molecular Biology |
| Pages | 63-80 |
| Number of pages | 18 |
| Volume | 783 |
| DOIs | |
| State | Published - 2011 |
Publication series
| Name | Methods in Molecular Biology |
|---|---|
| Volume | 783 |
| ISSN (Print) | 10643745 |
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Keywords
- Budding yeast
- Cytoplasmic dynein
- Microtubules
- Molecular motors
- Optical trapping
- Optical tweezers
- Saccharomyces cerevisiae
- Single-molecule assays
ASJC Scopus subject areas
- Molecular Biology
- Genetics
Cite this
Probing the force generation and stepping behavior of cytoplasmic dynein. / Gennerich, Arne; Reck-Peterson, Samara L.
Methods in Molecular Biology. Vol. 783 2011. p. 63-80 (Methods in Molecular Biology; Vol. 783).Research output: Chapter in Book/Report/Conference proceeding › Chapter
}
TY - CHAP
T1 - Probing the force generation and stepping behavior of cytoplasmic dynein
AU - Gennerich, Arne
AU - Reck-Peterson, Samara L.
PY - 2011
Y1 - 2011
N2 - Cytoplasmic dynein, which is the largest and arguably the most complex cytoskeletal motor protein, plays fundamental roles during cell division, nuclear positioning, and organelle and mRNA transport, by generating force and movement toward the minus ends of microtubules. Consequently, dynein is central to many physiological processes, and its dysfunction is implicated in human diseases. However, the molecular mechanism by which dynein produces force and movement remains poorly understood. Here, we describe the use of optical tweezers to probe the nanometer-scale motion and force generation of individual dynein molecules, and provide a hands-on protocol for how to purify cytoplasmic dynein from budding yeast in amounts sufficient for single-molecule studies.
AB - Cytoplasmic dynein, which is the largest and arguably the most complex cytoskeletal motor protein, plays fundamental roles during cell division, nuclear positioning, and organelle and mRNA transport, by generating force and movement toward the minus ends of microtubules. Consequently, dynein is central to many physiological processes, and its dysfunction is implicated in human diseases. However, the molecular mechanism by which dynein produces force and movement remains poorly understood. Here, we describe the use of optical tweezers to probe the nanometer-scale motion and force generation of individual dynein molecules, and provide a hands-on protocol for how to purify cytoplasmic dynein from budding yeast in amounts sufficient for single-molecule studies.
KW - Budding yeast
KW - Cytoplasmic dynein
KW - Microtubules
KW - Molecular motors
KW - Optical trapping
KW - Optical tweezers
KW - Saccharomyces cerevisiae
KW - Single-molecule assays
UR - http://www.scopus.com/inward/record.url?scp=80054756985&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=80054756985&partnerID=8YFLogxK
U2 - 10.1007/978-1-61779-282-3_4
DO - 10.1007/978-1-61779-282-3_4
M3 - Chapter
SN - 9781617792816
VL - 783
T3 - Methods in Molecular Biology
SP - 63
EP - 80
BT - Methods in Molecular Biology
ER -