TY - CHAP
T1 - Probing the force generation and stepping behavior of cytoplasmic dynein
AU - Gennerich, Arne
AU - Reck-Peterson, Samara L.
PY - 2011/10/24
Y1 - 2011/10/24
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
C2 - 21909883
AN - SCOPUS:80054756985
SN - 9781617792816
T3 - Methods in Molecular Biology
SP - 63
EP - 80
BT - Single Molecule Analysis
A2 - Peterman, Erwin J.G.
A2 - Wuite, Gijs J.L.
ER -