Name: Single-molecule FRET-beeldvorming voor het observeren van de conformationele dynamiek van dynamin-achtig GTPase-atlastine (Video) | JoVE Journal
Uploaded: 2025-01-24T13:40:00.000+00:00
Duration: 10 min 19 s
Description: Leer single-molecule FRET-beeldvorming voor het observeren van de conformationele dynamiek van atlastine tijdens de GTP-hydrolyse. Deze methode biedt inzicht in membraanfusieprocessen.

ai_agent_IP

ai_agent

maintenance

instantTrialDurationMinutes

isBiopharma

enableNewMyJoveSectionDesign

educationFeatures

seoMetaApi

ai_copilot_institution_blacklist

ai_chat_institution_blacklist

freeAccessWindow

ssoRoleCapture

scheduledMaintenanceBanner

joveLabsFeatures

disable_autoplay_for_bots

extended_trial_duration

5 minutes

<meta charset="utf8"></head><body>Monitoring van GTPase-atlastine in verschillende nucleotidelaadtoestanden

<ol><li>Jimah, J. R., Hinshaw, J. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Structural+insights+into+the+mechanism+of+dynamin+superfamily+proteins%5BTitle%5D)+AND+%22Trends+Cell+Biol%22%5BJournal%5D)">Structural insights into the mechanism of dynamin superfamily proteins.</a> Trends Cell Biol. 29 (3), 257-273 (2019).</li>
<li>Kalia, R., Frost, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Open+and+cut%3A+Allosteric+motion+and+membrane+fission+by+dynamin+superfamily+proteins%5BTitle%5D)+AND+%22Mol+Biol+Cell%22%5BJournal%5D)">Open and cut: Allosteric motion and membrane fission by dynamin superfamily proteins.</a> Mol Biol Cell. 30 (17), 2097-2104 (2019).</li>
<li>Ramachandran, R., Schmid, S. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+dynamin+superfamily%5BTitle%5D)+AND+%22Curr+Biol%22%5BJournal%5D)">The dynamin superfamily.</a> Curr Biol. 28 (8), R411-R416 (2018).</li>
<li>Bryce, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Human+atlastin-3+is+a+constitutive+ER+membrane+fusion+catalyst%5BTitle%5D)+AND+%22J+Cell+Biol%22%5BJournal%5D)">Human atlastin-3 is a constitutive ER membrane fusion catalyst.</a> J Cell Biol. 222 (7), e202211021(2023).</li>
<li>Crosby, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Reconstitution+of+human+atlastin+fusion+activity+reveals+autoinhibition+by+the+C+terminus%5BTitle%5D)+AND+%22J+Cell+Biol%22%5BJournal%5D)">Reconstitution of human atlastin fusion activity reveals autoinhibition by the C terminus.</a> J Cell Biol. 221 (2), e202107070(2022).</li>
<li>Hu, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+class+of+dynamin-like+GTPases+involved+in+the+generation+of+the+tubular+er+network%5BTitle%5D)+AND+%22Cell%22%5BJournal%5D)">A class of dynamin-like GTPases involved in the generation of the tubular er network.</a> Cell. 138 (3), 549-561 (2009).</li>
<li>Jang, E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Human+atlastins+are+sufficient+to+drive+the+fusion+of+liposomes+with+a+physiological+lipid+composition%5BTitle%5D)+AND+%22J+Cell+Biol%22%5BJournal%5D)">Human atlastins are sufficient to drive the fusion of liposomes with a physiological lipid composition.</a> J Cell Biol. 222 (4), e202109090(2023).</li>
<li>Liu, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Atlastin-1+regulates+morphology+and+function+of+endoplasmic+reticulum+in+dendrites%5BTitle%5D)+AND+%22Nat+Commun%22%5BJournal%5D)">Atlastin-1 regulates morphology and function of endoplasmic reticulum in dendrites.</a> Nat Commun. 10 (1), 568(2019).</li>
<li>Orso, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Homotypic+fusion+of+ER+membranes+requires+the+dynamin-like+gtpase+atlastin%5BTitle%5D)+AND+%22Nature%22%5BJournal%5D)">Homotypic fusion of ER membranes requires the dynamin-like gtpase atlastin.</a> Nature. 460 (7258), 978-983 (2009).</li>
<li>Rismanchi, N., Soderblom, C., Stadler, J., Zhu, P. P., Blackstone, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Atlastin+GTPases+are+required+for+Golgi+apparatus+and+er+morphogenesis%5BTitle%5D)+AND+%22Hum+Mol+Genet%22%5BJournal%5D)">Atlastin GTPases are required for Golgi apparatus and er morphogenesis.</a> Hum Mol Genet. 17 (11), 1591-1604 (2008).</li>
<li>Faust, J. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+atlastin+C-terminal+tail+is+an+amphipathic+helix+that+perturbs+the+bilayer+structure+during+endoplasmic+reticulum+homotypic+fusion%5BTitle%5D)+AND+%22J+Biol+Chem%22%5BJournal%5D)">The atlastin C-terminal tail is an amphipathic helix that perturbs the bilayer structure during endoplasmic reticulum homotypic fusion.</a> J Biol Chem. 290 (8), 4772-4783 (2015).</li>
<li>Liu, T. Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Lipid+interaction+of+the+c+terminus+and+association+of+the+transmembrane+segments+facilitate+atlastin-mediated+homotypic+endoplasmic+reticulum+fusion%5BTitle%5D)+AND+%22Proc+Natl+Acad+Sci+U+S+A%22%5BJournal%5D)">Lipid interaction of the c terminus and association of the transmembrane segments facilitate atlastin-mediated homotypic endoplasmic reticulum fusion.</a> Proc Natl Acad Sci U S A. 109 (32), E2146-E2154 (2012).</li>
<li>Moss, T. J., Andreazza, C., Verma, A., Daga, A., Mcnew, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Membrane+fusion+by+the+GTPase+atlastin+requires+a+conserved+c-terminal+cytoplasmic+tail+and+dimerization+through+the+middle+domain%5BTitle%5D)+AND+%22Proc+Natl+Acad+Sci+U+S+A%22%5BJournal%5D)">Membrane fusion by the GTPase atlastin requires a conserved c-terminal cytoplasmic tail and dimerization through the middle domain.</a> Proc Natl Acad Sci U S A. 108 (27), 11133-11138 (2011).</li>
<li>Bian, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Structures+of+the+atlastin+GTpase+provide+insight+into+homotypic+fusion+of+endoplasmic+reticulum+membranes%5BTitle%5D)+AND+%22Proc+Natl+Acad+Sci+U+S+A%22%5BJournal%5D)">Structures of the atlastin GTpase provide insight into homotypic fusion of endoplasmic reticulum membranes.</a> Proc Natl Acad Sci U S A. 108 (10), 3976-3981 (2011).</li>
<li>Byrnes, L. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Structural+basis+for+conformational+switching+and+GTP+loading+of+the+large+g+protein+atlastin%5BTitle%5D)+AND+%22EMBO+J%22%5BJournal%5D)">Structural basis for conformational switching and GTP loading of the large g protein atlastin.</a> EMBO J. 32 (3), 369-384 (2013).</li>
<li>Byrnes, L. J., Sondermann, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Structural+basis+for+the+nucleotide-dependent+dimerization+of+the+large+g+protein+atlastin-1%2Fspg3a%5BTitle%5D)+AND+%22Proc+Natl+Acad+Sci+U+S+A%22%5BJournal%5D)">Structural basis for the nucleotide-dependent dimerization of the large g protein atlastin-1/spg3a.</a> Proc Natl Acad Sci U S A. 108 (6), 2216-2221 (2011).</li>
<li>Morin-Leisk, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((An+intramolecular+salt+bridge+drives+the+soluble+domain+of+gtp-bound+atlastin+into+the+postfusion+conformation%5BTitle%5D)+AND+%22J+Cell+Biol%22%5BJournal%5D)">An intramolecular salt bridge drives the soluble domain of gtp-bound atlastin into the postfusion conformation.</a> J Cell Biol. 195 (4), 605-615 (2011).</li>
<li>Pendin, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((GTP-dependent+packing+of+a+three-helix+bundle+is+required+for+atlastin-mediated+fusion%5BTitle%5D)+AND+%22Proc+Natl+Acad+Sci+U+S+A%22%5BJournal%5D)">GTP-dependent packing of a three-helix bundle is required for atlastin-mediated fusion.</a> Proc Natl Acad Sci U S A. 108 (39), 16283-16288 (2011).</li>
<li>Winsor, J., Hackney, D. D., Lee, T. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+crossover+conformational+shift+of+the+GTPase+atlastin+provides+the+energy+driving+ER+fusion%5BTitle%5D)+AND+%22J+Cell+Biol%22%5BJournal%5D)">The crossover conformational shift of the GTPase atlastin provides the energy driving ER fusion.</a> J Cell Biol. 216 (5), 1321-1335 (2017).</li>
<li>Yan, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Structures+of+the+yeast+dynamin-like+GTPase+sey1p+provide+insight+into+homotypic+er+fusion%5BTitle%5D)+AND+%22J+Cell+Biol%22%5BJournal%5D)">Structures of the yeast dynamin-like GTPase sey1p provide insight into homotypic er fusion.</a> J Cell Biol. 210 (6), 961-972 (2015).</li>
<li>Rizo, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Molecular+mechanisms+underlying+neurotransmitter+release%5BTitle%5D)+AND+%22Annu+Rev+Biophys%22%5BJournal%5D)">Molecular mechanisms underlying neurotransmitter release.</a> Annu Rev Biophys. 51, 377-408 (2022).</li>
<li>Gao, S., Hu, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mitochondrial+fusion%3A+The+machineries+in+and+out%5BTitle%5D)+AND+%22Trends+Cell+Biol%22%5BJournal%5D)">Mitochondrial fusion: The machineries in and out.</a> Trends Cell Biol. 31 (1), 62-74 (2021).</li>
<li>Kelly, C. M., Byrnes, L. J., Neela, N., Sondermann, H., O'donnell, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+hypervariable+region+of+atlastin-1+is+a+site+for+intrinsic+and+extrinsic+regulation%5BTitle%5D)+AND+%22J+Cell+Biol%22%5BJournal%5D)">The hypervariable region of atlastin-1 is a site for intrinsic and extrinsic regulation.</a> J Cell Biol. 220 (11), e202104128(2021).</li>
<li>Asher, W. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((GPCR-mediated+beta-arrestin+activation+deconvoluted+with+single-molecule+precision%5BTitle%5D)+AND+%22Cell%22%5BJournal%5D)">GPCR-mediated beta-arrestin activation deconvoluted with single-molecule precision.</a> Cell. 185 (10), 1661-1675.e16 (2022).</li>
<li>Li, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mechanism+of+DNA+translocation+underlying+chromatin+remodelling+by+snf2%5BTitle%5D)+AND+%22Nature%22%5BJournal%5D)">Mechanism of DNA translocation underlying chromatin remodelling by snf2.</a> Nature. 567 (7748), 409-413 (2019).</li>
<li>Chen, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Conformational+dynamics+of+dynamin-like+MXA+revealed+by+single-molecule+fret%5BTitle%5D)+AND+%22Nat+Commun%22%5BJournal%5D)">Conformational dynamics of dynamin-like MXA revealed by single-molecule fret.</a> Nat Commun. 8, 15744(2017).</li>
<li>Shi, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Dissecting+the+mechanism+of+atlastin-mediated+homotypic+membrane+fusion+at+the+single-molecule+level%5BTitle%5D)+AND+%22Nat+Commun%22%5BJournal%5D)">Dissecting the mechanism of atlastin-mediated homotypic membrane fusion at the single-molecule level.</a> Nat Commun. 15 (1), 2488(2024).</li>
<li>Guelly, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Targeted+high-throughput+sequencing+identifies+mutations+in+atlastin-1+as+a+cause+of+hereditary+sensory+neuropathy+type+i%5BTitle%5D)+AND+%22Am+J+Hum+Genet%22%5BJournal%5D)">Targeted high-throughput sequencing identifies mutations in atlastin-1 as a cause of hereditary sensory neuropathy type i.</a> Am J Hum Genet. 88 (1), 99-105 (2011).</li>
<li>Montagna, A., Vajente, N., Pendin, D., Daga, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((In+vivo+analysis+of+CRISPR%2FCas9+induced+atlastin+pathological+mutations+in+drosophila%5BTitle%5D)+AND+%22Front+Neurosci%22%5BJournal%5D)">In vivo analysis of CRISPR/Cas9 induced atlastin pathological mutations in drosophila.</a> Front Neurosci. 14, 547746(2020).</li>
<li>Ulengin, I., Park, J. J., Lee, T. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((ER+network+formation+and+membrane+fusion+by+atlastin1%2Fspg3a+disease+variants%5BTitle%5D)+AND+%22Mol+Biol+Cell%22%5BJournal%5D)">ER network formation and membrane fusion by atlastin1/spg3a disease variants.</a> Mol Biol Cell. 26 (9), 1616-1628 (2015).</li>
<li>Zhao, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mutations+in+a+newly+identified+GTPase+gene+cause+autosomal+dominant+hereditary+spastic+paraplegia%5BTitle%5D)+AND+%22Nat+Genet%22%5BJournal%5D)">Mutations in a newly identified GTPase gene cause autosomal dominant hereditary spastic paraplegia.</a> Nat Genet. 29 (3), 326-331 (2001).</li>
</ol>

<table><tbody><tr><td>10 K Centrifugal Filters</td><td>Amicon</td><td>UFC901096</td><td></td></tr><tr><td>3-Aminopropyltriethoxysilaan (APTES)&nbsp;</td><td>Sigma-Aldrich</td><td>440140</td><td></td></tr><tr><td>45 Ti rotor</td><td></td><td></td><td></td></tr><tr><td>532-nm laser</td><td>Olympus</td><td></td><td></td></tr><tr><td>640-nm laser</td><td>Olympus</td><td></td><td></td></tr><tr><td>7 K MWCO spin desalting columns</td><td>Thermo Scientific</td><td>89882</td><td></td></tr><tr><td>Adenosine 5-trifosfaat dinatriumzout (ATP)</td><td>Roche</td><td>11140965001</td><td></td></tr><tr><td>Benzoëzuur</td><td>Sigma-Aldrich</td><td>242381</td><td></td></tr><tr><td>Biotine-PEG-SVA-5000</td><td>Laysan Bio</td><td>170-124</td><td></td></tr><tr><td>Gebiotinileerd anti His-antilichaam</td><td>Bioss Antibodies</td><td>bs-0287R-bio</td><td></td></tr><tr><td>d-biotine</td><td>Sigma-Aldrich</td><td>B4501</td><td></td></tr><tr><td>EDTA-vrije proteaseremmercocktail</td><td>Roche</td><td>11873580001</td><td></td></tr><tr><td>EMCCD-camera</td><td>Andor</td><td>IX897</td><td></td></tr><tr><td>Guanosine 5'-difosfaat natriumzout (GDP)</td><td>Sigma-Aldrich</td><td>&nbsp;G7127</td><td></td></tr><tr><td>Guanosine 5'-O-[gamma-thio]trifosfaat (GTP'S)</td><td>Roche</td><td>10220647001</td><td></td></tr><tr><td>Olie-immersieobjectief met hoge numerieke apertuur</td><td>Nikon</td><td></td><td>Nikon, 100x, N.A. 1.49, oliejmersie</td></tr><tr><td>ImageJ</td><td>NIH</td><td></td><td>https://imagej.net/ij/</td></tr><tr><td>Isopropyl-β-D-thiogalactosid</td><td>Sigma-Aldrich</td><td>I5502</td><td></td></tr><tr><td>LD555-MAL</td><td>Lumidyne</td><td>4</td><td></td></tr><tr><td>LD655-MAL</td><td>Lumidyne</td><td>10</td><td></td></tr><tr><td>L-glutaminezuurkalium</td><td>Sigma-Aldrich</td><td>G1501</td><td></td></tr><tr><td>Magnesiumacetaattetrahydraat</td><td>Sigma-Aldrich</td><td>M0631</td><td></td></tr><tr><td>Matlab</td><td>The MathWorks, Natick, MA</td><td></td><td>https://www.mathworks.com/</td></tr><tr><td>Microscoopdekglas</td><td>Fisherbrand</td><td>18834</td><td>&nbsp;(24 mm &times; 60 mm)</td></tr><tr><td>Microscoopglaasjes</td><td></td><td></td><td>Aangepast, (75 mm &times; 26 mm &times; 1 mm) met 6 paren doorgaande gaten met een diameter van 1,2 mm</td></tr><tr><td>mPEG-SVA-5000</td><td>Laysan Bio</td><td>170-106</td><td></td></tr><tr><td>Ni Sepharose 6 Fast Flow</td><td>Cytiva</td><td>17531802</td><td></td></tr><tr><td>Origin</td><td>Origin software</td><td></td><td>https://www.originlab.com/</td></tr><tr><td>Polystyreen-deeltjes</td><td>QDSphere</td><td>AG1265</td><td></td></tr><tr><td>Protocatechuaat 3,4-dioxygeenase</td><td>Sigma-Aldrich</td><td>P8279</td><td></td></tr><tr><td>Streptavidine</td><td>Sangon Biotech</td><td>A610492</td><td></td></tr><tr><td>Superdex 200 Increase (10/300 GL)</td><td>Cytiva</td><td>28990944</td><td></td></tr><tr><td>TIRF-microscoop</td><td>Nikon</td><td>Ti2</td><td></td></tr><tr><td>Tris(2-carboxyethyl)fosfinehydrochloride (TCEP)</td><td>Thermo</td><td>75259</td><td></td></tr></tbody></table>

single-molecule fret-beeldvorming voor het observeren van de conformationele dynamiek van dynamin-achtig gtpase-atlastine

Single-Molecule FRET Imaging for Observing the Conformational Dynamics of Dynamin-Like GTPase Atlastin

Single-molecule FRET-beeldvorming voor het observeren van de conformationele dynamiek van dynamin-achtig GTPase-atlastine

The rigid-body rotation of the three-helical middle domain (3HB) relative to the GTPase domain of dynamin-like protein atlastin (ATL) is a crucial driver ...

single-molecule-fret-imaging-for-observing-conformational-dynamics

Research

JoVE Journal

Biochemistry

Biochemie

This video describes TIRF, a total internal reflection microscopy-based technique to visualize actin and microtubule polymerization dynamics. The method allows the high-resolution imaging of actin and microtubule coupling dynamics in real-time, which is essential for understanding cellular crosstalks.

TIRF Microscopy to Visualize Actin and Microtubule Coupling Dynamics

Single fluoroforen kunnen worden gelokaliseerd met nanometer precisie met behulp van FIONA. Hier een overzicht van de FIONA techniek is gemeld, en hoe FIONA experimenten uit te voeren wordt beschreven.

Fluorescentie Imaging met One-nanometer nauwkeurigheid (FIONA)

Hier we illustreren hoe enkel molecuul foto-geactiveerde lokalisatie microscopie kan worden uitgevoerd op de motorische zenuw-terminal van een levende Drosophila melanogaster derde instar larve.

In Vivo Single-molecuul volgen bij de Terminal van de presynaptische motorische zenuw Drosophila

We beschrijven hoe micro- en photomanipulation technieken zoals FRAP en photoactivation inschakelen de bepaling van de parameters van de beweeglijkheid en de spatio dynamiek van eiwitten in het migreren van cellen. Experimentele uitlezingen omvatten subcellular dynamiek en omzet van beweeglijkheid toezichthouders of van de onderliggende actine cytoskelet.

Micromanipulation technieken waardoor analyse van morfogenetische dynamiek en een omzet van Cytoskeletal regelgevers

Hier presenteren we een protocol voor het gelijktijdig gebruik van Förster resonance energie overdracht gebaseerde spanning sensoren voor het meten van eiwit belasting en fluorescentie herstel na photobleaching voor het meten van de dynamiek van de eiwitten waardoor de meting van kracht-gevoelige eiwit dynamiek binnen levende cellen.

Meting van de kracht-gevoelige eiwitten Dynamics in levende cellen met behulp van een combinatie van Fluorescent technieken

We presenteren protocollen voor eenvoudige actinefilamentmicrofluïdische assays, in combinatie met fluorescentiemicroscopie, waarmee men individuele actinefilamenten in realtime nauwkeurig kan volgen terwijl ze sequentieel worden blootgesteld aan verschillende eiwitoplossingen.

Microfluïdica en fluorescentiemicroscopie gebruiken om de assemblagedynamiek van single actinefilamenten en -bundels te bestuderen

Dit protocol beschrijft de vorming van ondersteunde lipide bilayers en de toevoeging van cytoskeletale filamenten en motoreiwitten om de dynamiek van gereconstitueerde, membraangebonden cytoskeletale netwerken te bestuderen met behulp van fluorescentiemicroscopie.

Reconstitutie van membraangebonden minimale actinecorticosterces op ondersteunde lipidebilagen

Dit protocol is een gids voor het visualiseren van dynamische actine en microtubuli met behulp van een in vitro total internal fluorescence (TIRF) microscopie assay.

Visualisatie van actine en microtubulikoppelingsdynamiek in vitro door total internal reflection fluorescence (TIRF) microscopie

Dit protocol beschrijft hoe TIRF-microscopie kan worden gebruikt om individuele ionkanalen te volgen en hun activiteit in ondersteunde lipidemembranen te bepalen, waardoor het samenspel tussen laterale membraanbeweging en kanaalfunctie wordt gedefinieerd. Het beschrijft hoe de membranen moeten worden voorbereid, de gegevens moeten worden vastgelegd en de resultaten moeten worden geanalyseerd.

Single-Molecule Imaging van Laterale Mobiliteit en Ion Channel Activity in Lipid Bilayers met behulp van Total Internal Reflection Fluorescence (TIRF) Microscopie

We rapporteren een hybride ensemble- en single-molecule-assay om de beweging van fluorescerend gelabelde, volledig gereconstitueerde Cdc45/Mcm2-7/GINS (CMG) helicase op lineaire DNA-moleculen die op hun plaats worden gehouden in een optische val, direct in beeld te brengen en te kwantificeren.

Hybride ensemble en single-molecule assay om de beweging van volledig gereconstitueerde CMG in beeld te brengen

Hier presenteren we methoden voor het bereiden van quasi-tweedimensionale (2D) verstrengelde, verknoopte en vloeibare kristalactine-assemblages van gezuiverde eiwitten.

Het afstemmen van de contractiliteit en vervormingsmodi van actieve op actine gebaseerde assemblages in vitro: van tweedimensionale actieve netwerken tot druppels van vloeibare kristallen

Leer single-molecule FRET-beeldvorming voor het observeren van de conformationele dynamiek van atlastine tijdens de GTP-hydrolyse. Deze methode biedt inzicht in membraanfusieprocessen.

Single-molecule FRET-beeldvorming voor het observeren van de conformationele dynamiek van dynamin-achtig GTPase-atlastine

Meer video's verkennen