Individual dimers of the mitotic kinesin motor Eg5 step processively and support substantial loads in vitro
Eg5, a member of the kinesin superfamily of microtubule-based motors, is essential for bipolar spindle assembly and maintenance during mitosis, yet little is known about the mechanisms by which it accomplishes these tasks. Here, we used an automated optical trapping apparatus in conjunction with a n...
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| Veröffentlicht in: | Nature cell biology 2006-05, Vol.8 (5), p.470-476 |
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| creator | Valentine, Megan T. Fordyce, Polly M. Krzysiak, Troy C. Gilbert, Susan P. Block, Steven M. |
| description | Eg5, a member of the kinesin superfamily of microtubule-based motors, is essential for bipolar spindle assembly and maintenance during mitosis, yet little is known about the mechanisms by which it accomplishes these tasks. Here, we used an automated optical trapping apparatus in conjunction with a novel motility assay that employed chemically modified surfaces to probe the mechanochemistry of Eg5. Individual dimers, formed by a recombinant human construct Eg5–513–5His, stepped processively along microtubules in 8-nm increments, with short run lengths averaging approximately eight steps. By varying the applied load (with a force clamp) and the ATP concentration, we found that the velocity of Eg5 was slower and less sensitive to external load than that of conventional kinesin, possibly reflecting the distinct demands of spindle assembly as compared with vesicle transport. The Eg5–513–5His velocity data were described by a minimal, three-state model where a force-dependent transition follows nucleotide binding. |
| doi_str_mv | 10.1038/ncb1394 |
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The Eg5–513–5His velocity data were described by a minimal, three-state model where a force-dependent transition follows nucleotide binding.</description><subject>Adenosine Triphosphate - metabolism</subject><subject>Animals</subject><subject>Assaying</subject><subject>ATP</subject><subject>Biomechanical Phenomena</subject><subject>Biomedical and Life Sciences</subject><subject>Cancer Research</subject><subject>Cattle</subject><subject>Cell Biology</subject><subject>Chemical properties</subject><subject>Developmental Biology</subject><subject>Dimerization</subject><subject>Histidine</subject><subject>Humans</subject><subject>Kinesin</subject><subject>Kinesin - chemistry</subject><subject>Kinesin - metabolism</subject><subject>Kinetics</subject><subject>letter</subject><subject>Life Sciences</subject><subject>Mechanical chemistry</subject><subject>Mechanical properties</subject><subject>Methods</subject><subject>Mitosis</subject><subject>Molecular Motor Proteins - chemistry</subject><subject>Molecular Motor Proteins - metabolism</subject><subject>Motility</subject><subject>Oligopeptides</subject><subject>Physiological aspects</subject><subject>Protein Structure, Quaternary</subject><subject>Recombinant Fusion Proteins - metabolism</subject><subject>Stem Cells</subject><subject>Trapping</subject><subject>Velocity</subject><issn>1465-7392</issn><issn>1476-4679</issn><issn>1476-4679</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2006</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><recordid>eNqFkk1v1DAQhiMEoqUg_gGyOEA5pNiJP-ILUlWVslIlJD7OlmNPtm4TO9jOiv57vNoVZXvBlxl5nnntdzRV9ZrgM4Lb7qM3PWklfVIdEyp4TbmQT7c5Z7VoZXNUvUjpFmNCKRbPqyPCOaaYs-PqbuWt2zi76BFZN0FMKAwo3wCaXA7ZGXTnPCTn0RRyiOhyzVDKMKM5BgMpuQ2M90h7i9IyzyHmEvuUtc-uKI5B24RK88blGF5WzwY9Jni1jyfVz8-XPy6-1Ndfr1YX59e1YVLkuu847S3DlGApmaXWaKOhs7wRVnSEQUvbtikZ7yijxaIpvpquH0BKTHrdnlSfdrrz0k9gDfgc9ajm6CYd71XQTh1WvLtR67BRhDVtS2gReLcXiOHXAimrySUD46g9hCWpMl0my-P_BYkgUjSYFPDtI_A2LNGXKaimHIk7wQt0toPWegTl_BDK77bmLUzOBA-DK_fnpGsx64QUpeHDQUNhMvzOa72kpFbfvx2y73esiSGlCMPfeRCstjuk9jtUyDf_ju-B2y9NAU53QColv4b4YOax1h8LDc79</recordid><startdate>20060501</startdate><enddate>20060501</enddate><creator>Valentine, Megan T.</creator><creator>Fordyce, Polly M.</creator><creator>Krzysiak, Troy C.</creator><creator>Gilbert, Susan P.</creator><creator>Block, Steven M.</creator><general>Nature Publishing Group UK</general><general>Nature Publishing Group</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>ISR</scope><scope>3V.</scope><scope>7QL</scope><scope>7QP</scope><scope>7QR</scope><scope>7T5</scope><scope>7TK</scope><scope>7TM</scope><scope>7TO</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>8AO</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20060501</creationdate><title>Individual dimers of the mitotic kinesin motor Eg5 step processively and support substantial loads in vitro</title><author>Valentine, Megan T. ; Fordyce, Polly M. ; Krzysiak, Troy C. ; Gilbert, Susan P. ; Block, Steven M.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c597t-b864bd50410995d4dcacae8d627d7815e3433278168454465c01428bfe9901ba3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2006</creationdate><topic>Adenosine Triphosphate - 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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Nature cell biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Valentine, Megan T.</au><au>Fordyce, Polly M.</au><au>Krzysiak, Troy C.</au><au>Gilbert, Susan P.</au><au>Block, Steven M.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Individual dimers of the mitotic kinesin motor Eg5 step processively and support substantial loads in vitro</atitle><jtitle>Nature cell biology</jtitle><stitle>Nat Cell Biol</stitle><addtitle>Nat Cell Biol</addtitle><date>2006-05-01</date><risdate>2006</risdate><volume>8</volume><issue>5</issue><spage>470</spage><epage>476</epage><pages>470-476</pages><issn>1465-7392</issn><issn>1476-4679</issn><eissn>1476-4679</eissn><abstract>Eg5, a member of the kinesin superfamily of microtubule-based motors, is essential for bipolar spindle assembly and maintenance during mitosis, yet little is known about the mechanisms by which it accomplishes these tasks. 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| subjects | Adenosine Triphosphate - metabolism Animals Assaying ATP Biomechanical Phenomena Biomedical and Life Sciences Cancer Research Cattle Cell Biology Chemical properties Developmental Biology Dimerization Histidine Humans Kinesin Kinesin - chemistry Kinesin - metabolism Kinetics letter Life Sciences Mechanical chemistry Mechanical properties Methods Mitosis Molecular Motor Proteins - chemistry Molecular Motor Proteins - metabolism Motility Oligopeptides Physiological aspects Protein Structure, Quaternary Recombinant Fusion Proteins - metabolism Stem Cells Trapping Velocity |
| title | Individual dimers of the mitotic kinesin motor Eg5 step processively and support substantial loads in vitro |
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