Name: Split-luciferase herassemblagetest om endoplasmatisch reticulum-mitochondriëncontacten in levende cellen te meten (Video) | JoVE Journal
Uploaded: 2024-10-11T13:10:00.000+00:00
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Description: Leer een split-luciferase herassemblagetest om endoplasmatisch reticulum-mitochondriëncontacten in levende HEK293T-cellen te meten. Deze methode vereenvoudigt de kwantificering van inter-organel communicatie voor onderzoek naar neurodegeneratie.

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<meta charset="utf8"></head><body>Een eenvoudige test om ER-mitochondriëncontacten te meten voor schermen met een hoge doorvoer

<ol><li>Aoyama-Ishiwatari, S., Hirabayashi, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Endoplasmic+reticulum-mitochondria+contact+sites-emerging+intracellular+signaling+hubs%5BTitle%5D)+AND+%22Front+Cell+Dev+Biol%22%5BJournal%5D)">Endoplasmic reticulum-mitochondria contact sites-emerging intracellular signaling hubs.</a> Front Cell Dev Biol. 9, 653828(2021).</li>
<li>Sassano, M. L., Felipe-Abrio, B., Agostinis, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((ER-mitochondria+contact+sites%3B+a+multifaceted+factory+for+Ca2%2B+signaling+and+lipid+transport%5BTitle%5D)+AND+%22Front+Cell+Dev+Biol%22%5BJournal%5D)">ER-mitochondria contact sites; a multifaceted factory for Ca2+ signaling and lipid transport.</a> Front Cell Dev Biol. 10, 988014(2022).</li>
<li>Scorrano, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Coming+together+to+define+membrane+contact+sites%5BTitle%5D)+AND+%22Nat+Commun%22%5BJournal%5D)">Coming together to define membrane contact sites.</a> Nat Commun. 10 (1), 1287(2019).</li>
<li>Voeltz, G. K., Sawyer, E. M., Hajnóczky, G., Prinz, W. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Making+the+connection%3A+How+membrane+contact+sites+have+changed+our+view+of+organelle+biology%5BTitle%5D)+AND+%22Cell%22%5BJournal%5D)">Making the connection: How membrane contact sites have changed our view of organelle biology.</a> Cell. 187 (2), 257-270 (2024).</li>
<li>Paillusson, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((There%27s+something+wrong+with+my+MAM%3B+the+ER-mitochondria+axis+and+neurodegenerative+diseases%5BTitle%5D)+AND+%22Trends+Neurosci%22%5BJournal%5D)">There's something wrong with my MAM; the ER-mitochondria axis and neurodegenerative diseases.</a> Trends Neurosci. 39 (3), 146-157 (2016).</li>
<li>Sasi, U. S. S., Sindhu, G., Raj, P. S., Raghu, K. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mitochondria+associated+membranes+%28MAMs%29%3A+emerging+drug+targets+for+diabetes%5BTitle%5D)+AND+%22Curr+Med+Chem%22%5BJournal%5D)">Mitochondria associated membranes (MAMs): emerging drug targets for diabetes.</a> Curr Med Chem. 27 (20), 3362-3385 (2019).</li>
<li>Joshi, A. U., Kornfeld, O. S., Mochly-Rosen, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+entangled+ER-mitochondrial+axis+as+a+potential+therapeutic+strategy+in+neurodegeneration%3A+A+tangled+duo+unchained%5BTitle%5D)+AND+%22Cell+Calcium%22%5BJournal%5D)">The entangled ER-mitochondrial axis as a potential therapeutic strategy in neurodegeneration: A tangled duo unchained.</a> Cell Calcium. 60 (3), 218-234 (2016).</li>
<li>Moltedo, O., Remondelli, P., Amodio, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+mitochondria-endoplasmic+reticulum+contacts+and+their+critical+role+in+aging+and+age-associated+diseases%5BTitle%5D)+AND+%22Front+Cell+Dev+Biol%22%5BJournal%5D)">The mitochondria-endoplasmic reticulum contacts and their critical role in aging and age-associated diseases.</a> Front Cell Dev Biol. 7, 172(2019).</li>
<li>Rieusset, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+role+of+endoplasmic+reticulum-mitochondria+contact+sites+in+the+control+of+glucose+homeostasis%3A+an+update%5BTitle%5D)+AND+%22Cell+Death+Dis%22%5BJournal%5D)">The role of endoplasmic reticulum-mitochondria contact sites in the control of glucose homeostasis: an update.</a> Cell Death Dis. 9 (3), 388(2018).</li>
<li>Bouguerra, M. D., Lalli, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((ER-mitochondria+interactions%3A+Both+strength+and+weakness+within+cancer+cells%5BTitle%5D)+AND+%22Biochim+Biophys+Acta+Mol+Cell+Res%22%5BJournal%5D)">ER-mitochondria interactions: Both strength and weakness within cancer cells.</a> Biochim Biophys Acta Mol Cell Res. 1866 (4), 650-662 (2019).</li>
<li>Tepikin, A. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mitochondrial+junctions+with+cellular+organelles%3A+Ca2%2B+signalling+perspective%5BTitle%5D)+AND+%22Pfl%C3%BCgers+Arch%22%5BJournal%5D)">Mitochondrial junctions with cellular organelles: Ca2+ signalling perspective.</a> Pflügers Arch. 470 (8), 1181-1192 (2018).</li>
<li>Masson, G. L., Przedborski, S., Abbott, L. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+computational+model+of+motor+neuron+degeneration%5BTitle%5D)+AND+%22Neuron%22%5BJournal%5D)">A computational model of motor neuron degeneration.</a> Neuron. 83 (4), 975-988 (2014).</li>
<li>Giamogante, F., Barazzuol, L., Brini, M., Calì, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((ER-mitochondria+contact+sites+reporters%3A+strengths+and+weaknesses+of+the+available+approaches%5BTitle%5D)+AND+%22Int+J+Mol+Sci%22%5BJournal%5D)">ER-mitochondria contact sites reporters: strengths and weaknesses of the available approaches.</a> Int J Mol Sci. 21 (21), 8157(2020).</li>
<li>Rizzuto, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Close+contacts+with+the+endoplasmic+reticulum+as+determinants+of+mitochondrial+Ca2%2B+responses%5BTitle%5D)+AND+%22Science%22%5BJournal%5D)">Close contacts with the endoplasmic reticulum as determinants of mitochondrial Ca2+ responses.</a> Science. 280 (5370), 1763-1766 (1998).</li>
<li>Valm, A. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Applying+systems-level+spectral+imaging+and+analysis+to+reveal+the+organelle+interactome%5BTitle%5D)+AND+%22Nature%22%5BJournal%5D)">Applying systems-level spectral imaging and analysis to reveal the organelle interactome.</a> Nature. 546 (7656), 162-167 (2017).</li>
<li>Csordás, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Imaging+interorganelle+contacts+and+local+calcium+dynamics+at+the+ER-mitochondrial+interface%5BTitle%5D)+AND+%22Mol+Cell%22%5BJournal%5D)">Imaging interorganelle contacts and local calcium dynamics at the ER-mitochondrial interface.</a> Mol Cell. 39 (1), 121-132 (2010).</li>
<li>Cieri, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((SPLICS%3A+a+split+green+fluorescent+protein-based+contact+site+sensor+for+narrow+and+wide+heterotypic+organelle+juxtaposition%5BTitle%5D)+AND+%22Cell+Death+Differ%22%5BJournal%5D)">SPLICS: a split green fluorescent protein-based contact site sensor for narrow and wide heterotypic organelle juxtaposition.</a> Cell Death Differ. 25 (6), 1131-1145 (2018).</li>
<li>Kakimoto, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Visualizing+multiple+inter-organelle+contact+sites+using+the+organelle-+targeted+split-GFP+system%5BTitle%5D)+AND+%22Sci+Rep%22%5BJournal%5D)">Visualizing multiple inter-organelle contact sites using the organelle- targeted split-GFP system.</a> Sci Rep. 8 (1), 6175(2018).</li>
<li>Wu, M. M., Covington, E. D., Lewis, R. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Single-molecule+analysis+of+diffusion+and+trapping+of+STIM1+and+Orai1+at+ER-plasma+membrane+junctions%5BTitle%5D)+AND+%22Mol+Biol+Cell%22%5BJournal%5D)">Single-molecule analysis of diffusion and trapping of STIM1 and Orai1 at ER-plasma membrane junctions.</a> Mol Biol Cell. 25 (22), Mac.E14-06-1107 (2014).</li>
<li>Csordas, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Structural+and+functional+features+and+significance+of+the+physical+linkage+between+ER+and+mitochondria%5BTitle%5D)+AND+%22J+Cell+Biol%22%5BJournal%5D)">Structural and functional features and significance of the physical linkage between ER and mitochondria.</a> J Cell Biol. 174 (7), 915-921 (2006).</li>
<li>de Brito, O. M., Scorrano, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mitofusin+2+tethers+endoplasmic+reticulum+to+mitochondria%5BTitle%5D)+AND+%22Nature%22%5BJournal%5D)">Mitofusin 2 tethers endoplasmic reticulum to mitochondria.</a> Nature. 456 (7222), 605-610 (2008).</li>
<li>Kremer, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Developing+3D+SEM+in+a+broad+biological+context%5BTitle%5D)+AND+%22J+Microsc%22%5BJournal%5D)">Developing 3D SEM in a broad biological context.</a> J Microsc. 259 (2), 80-96 (2015).</li>
<li>Söderberg, O., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Direct+observation+of+individual+endogenous+protein+complexes+in+situ+by+proximity+ligation%5BTitle%5D)+AND+%22Nat+Methods%22%5BJournal%5D)">Direct observation of individual endogenous protein complexes in situ by proximity ligation.</a> Nat Methods. 3 (12), 995-1000 (2006).</li>
<li>Lim, Y., Cho, I. -T., Rennke, H. G., Cho, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((%CE%B22-adrenergic+receptor+regulates+ER-mitochondria+contacts%5BTitle%5D)+AND+%22Sci+Rep%22%5BJournal%5D)">β2-adrenergic receptor regulates ER-mitochondria contacts.</a> Sci Rep. 11 (1), 21477(2021).</li>
<li>Lam, S. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Directed+evolution+of+APEX2+for+electron+microscopy+and+proximity+labeling%5BTitle%5D)+AND+%22Nat+Methods%22%5BJournal%5D)">Directed evolution of APEX2 for electron microscopy and proximity labeling.</a> Nat Methods. 12 (1), 51-54 (2014).</li>
<li>Cho, I. -T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Ascorbate+peroxidase+proximity+labeling+coupled+with+biochemical+fractionation+identifies+promoters+of+endoplasmic+reticulum-mitochondrial+contacts%5BTitle%5D)+AND+%22J+Biol+Chem%22%5BJournal%5D)">Ascorbate peroxidase proximity labeling coupled with biochemical fractionation identifies promoters of endoplasmic reticulum-mitochondrial contacts.</a> J Biol Chem. 292 (39), 16382-16392 (2017).</li>
<li>Lim, Y., Cho, I. -T., Schoel, L. J., Cho, G., Golden, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Hereditary+spastic+paraplegia-linked+REEP1+modulates+endoplasmic+reticulum%2Fmitochondria+contacts%5BTitle%5D)+AND+%22Ann+Neurol%22%5BJournal%5D)">Hereditary spastic paraplegia-linked REEP1 modulates endoplasmic reticulum/mitochondria contacts.</a> Ann Neurol. 78 (5), 679-696 (2015).</li>
<li>Arnold, T. R., Stephenson, R. E., Miller, A. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Rho+GTPases+and+actomyosin%3A+Partners+in+regulating+epithelial+cell-cell+junction+structure+and+function%5BTitle%5D)+AND+%22Exp+Cell+Res%22%5BJournal%5D)">Rho GTPases and actomyosin: Partners in regulating epithelial cell-cell junction structure and function.</a> Exp Cell Res. 358 (1), 20-30 (2017).</li>
</ol>

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split-luciferase herassemblagetest om endoplasmatisch reticulum-mitochondriëncontacten in levende cellen te meten

Split-Luciferase Reassembly Assay to Measure Endoplasmic Reticulum-Mitochondria Contacts in Live Cells

Split-luciferase herassemblagetest om endoplasmatisch reticulum-mitochondriëncontacten in levende cellen te meten

Endoplasmic reticulum (ER)-mitochondria contact sites play a critical role in cell health and homeostasis, such as the regulation of Ca2+ and lipid ...

split-luciferase-reassembly-assay-to-measure-endoplasmic-reticulum

Research

JoVE Journal

Biochemistry

Biochemie

Hier beschrijven we de procedures voor het bestuderen van veranderingen in de fagocytose-geïnduceerde genexpressie met een luciferase-based reportergen benadering met behulp van de Dual-GloTM Luciferase Assay System van Promega.

Studeren Membraan Biogenese met een Luciferase-Based Reporter Gene test

This video demonstrates the split luciferase complementation assay to detect protein-protein interactions. In this assay, the proteins of interest are tagged to small and large fragments of the luciferase enzyme. When the proteins interact, the large and small fragments combine to form an active enzyme complex, which in the presence of a specific substrate, releases bright luminescence that can be measured.

Split Luciferase Complementation Assay to Identify Specific Protein-Protein Interactions

Het bereiken van een systeemniveau begrip van cellulaire processen is een doel van de hedendaagse celbiologie. We beschrijven hier strategieën voor multiplexing luciferase reporters van verschillende cellulaire functie eindpunten naar gen-functie met behulp van genoom-schaal RNAi bibliotheken ondervragen.

Een Multiplexed Luciferase-gebaseerde screening Platform voor ondervragen Kanker-geassocieerde signaaltransductie in gekweekte cellen

Dit artikel beschrijft het gebruik van een vuurvlieg luciferase-GFP-fusie-eiwit te...

Gekoppeld Testen voor Monitoring Protein Refolding in<em&gt; Saccharomyces cerevisiae</em

We beschrijven beeldvormingsbenaderingen we gebruiken om de verdeling en mobiliteit van de getransfecteerde fluorescerende eiwitten in het endoplasmatisch reticulum (ER) inwoner via de confocale beeldvorming van levende cellen te onderzoeken. We hebben ook ultrastructureel analyseren het effect van hun expressie op de architectuur van deze subcellulaire compartiment.

Visualisatie van endoplasmatisch reticulum subdomeinen bij gekweekte cellen

Interacties tussen eiwitten zijn fundamenteel voor alle cellulaire processen. Met Bioluminescentie Resonance Energy Transfer, kan de interactie tussen twee eiwitten worden gecontroleerd in levende cellen en in real time. Bovendien kunnen de effecten van potentieel pathogene mutaties worden beoordeeld.

Onderzoeken eiwit-eiwit interacties in levende cellen met behulp van bioluminescentie resonantie energieoverbrenging

Hier beschrijven we een werkwijze voor het visualiseren en kwantificeren gevoeligheid een endogene interacties tussen het endoplasmatisch reticulum en mitochondria in gefixeerde cellen. Het protocol is voorzien van een geoptimaliseerde in situ nabijheid ligation assay gericht op de inositol 1,4,5-trifosfaat receptor / glucose-gereguleerd eiwit 75 / voltage-afhankelijke anion kanaal / cyclofiline D-complex aan de mitochondria bijbehorende membraan interface.

Studie van endoplasmatisch reticulum en mitochondriën interacties door<em&gt; In Situ</em&gt; Proximity Ligatie Assay in gefixeerde cellen

Het hier gepresenteerde protocol is bedoeld om het optreden van heterologe interacties tussen Golgi-residente type III membraaneiwitten met cytoplasmatisch blootgestelde N- en/of C-termini in levende zoogdiercellen aan te tonen met behulp van de meest recente variant van de split luciferase complementatie assay.

Demonstratie van heterologe complexen gevormd door Golgi-Resident Type III membraaneiwitten met behulp van Split Luciferase Complementation Assay

De behoefte aan nieuwe benaderingen voor het bestuderen van membraancontactplaatsen (MCS'en) is gegroeid als gevolg van de toenemende interesse in het bestuderen van deze cellulaire structuren en hun componenten. Hier presenteren we een protocol dat eerder beschikbare microscopietechnologieën integreert om intra-organel en inter-organel eiwitcomplexen die zich in MCS'en bevinden te identificeren en te kwantificeren.

Visualisatie en kwantificering van endogene intra-organeleiwitinteracties op ER-mitochondriale contactlocaties door nabijheidsligatietests

Dit protocol beschrijft een op BRET gebaseerde test voor het meten van de interacties van het CRAF-kinase met 14-3-3-eiwitten in levende cellen. Het protocol schetst stappen voor het voorbereiden van de cellen, het uitlezen van BRET-emissies en gegevensanalyse. Er wordt ook een voorbeeldresultaat gepresenteerd met identificatie van de juiste controles en probleemoplossing voor testoptimalisatie.

Op bioluminescentieresonantie energieoverdracht (BRET) gebaseerde test voor het meten van interacties van CRAF met 14-3-3-eiwitten in levende cellen

Leer een split-luciferase herassemblagetest om endoplasmatisch reticulum-mitochondriëncontacten in levende HEK293T-cellen te meten. Deze methode vereenvoudigt de kwantificering van inter-organel communicatie voor onderzoek naar neurodegeneratie.

Split-luciferase herassemblagetest om endoplasmatisch reticulum-mitochondriëncontacten in levende cellen te meten

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