Name: electrochemical detection of deuterium kinetic isotope effect on extracellular electron transport in shewanella oneidensis mr-1
Uploaded: 2018-04-16T14:30:00
Description: Watch this Scientific Journal Video about Electrochemical Detection of Deuterium Kinetic Isotope Effect on Extracellular Electron Transport in Shewanella oneidensis MR-1 at JoVE.com

Detta arbete stöddes ekonomiskt av ett bidrag för speciellt främjat forskning från Japan sällskapet för främjande av vetenskap (JSPS) KAKENHI licensnummer 24000010, 17H 04969, och JP17J02602, den oss Office av Naval Research globala (N62909-17-1-2038). Y.T. är JSPS forskarassistent och stöds av JSPS genom Program för ledande Graduate skolor (MERIT).

1. bildandet av en enskiktslager Biofilm av S. oneidensis MR-1 på en ITO elektrod (figur 1)
Obs: För att förhindra kontaminering av elektrokemiska reaktorn med andra mikrober, alla media, redskap, och komponenter av elektrokemiska reaktorn ska steriliseras i förväg. När du använder S. oneidensis MR-1 celler och konstruera de elektrokemiska reaktorerna, alla förfaranden bör utföras på en ren bänk.
<ol><li>Odling av S. oneidensis<...

<ol>
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Microbiol. 48, 311-343 (1994).</li><li>Bretschger, O., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Current+production+and+metal+oxide+reduction+by+Shewanella+oneidensis+MR-1+wild+type+and+mutants.">Current production and metal oxide reduction by Shewanella oneidensis MR-1 wild type and mutants.</a> Appl Environ Microb. 73 (21), 7003-7012 (2007).</li><li>Richter, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cyclic+voltammetry+of+biofilms+of+wild+type+and+mutant+Geobacter+sulfurreducens+on+fuel+cell+anodes+indicates+possible+roles+of+OmcB,+OmcZ,+type+IV+pili,+and+protons+in+extracellular+electron+transfer.">Cyclic voltammetry of biofilms of wild type and mutant Geobacter sulfurreducens on fuel cell anodes indicates possible roles of OmcB, OmcZ, type IV pili, and protons in extracellular electron transfer.</a> Energy Environ. 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Ed. 56, 9082-9086 (2017).</li><li>Hammes-Schiffer, S., Stuchebrukhov, A. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Theory+of+Coupled+Electron+and+Proton+Transfer+Reactions.">Theory of Coupled Electron and Proton Transfer Reactions.</a> Chem. Rev. 110 (12), 6939-6960 (2010).</li><li>Cleland, W. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+use+of+isotope+effects+to+determine+enzyme+mechanisms.">The use of isotope effects to determine enzyme mechanisms.</a> J Biol. Chem. 278 (52), 51975-51984 (2003).</li><li>Kouzuma, A., Kasai, T., Hirose, A., Watanabe, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Catabolic+and+regulatory+systems+in+Shewanella+oneidensis+MR-1+involved+in+electricity+generation+in+microbial+fuel+cells.">Catabolic and regulatory systems in Shewanella oneidensis MR-1 involved in electricity generation in microbial fuel cells.</a> Front. Microbiol. 6, (2015).</li><li>Kushner, D. J., Baker, A., Dunstall, T. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pharmacological+uses+and+perspectives+of+heavy+water+and+deuterated+compounds.">Pharmacological uses and perspectives of heavy water and deuterated compounds.</a> Can. J Physiol. Pharm. 77 (2), 79-88 (1999).</li><li>Xie, X. S., Zubarev, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+Low-Level+Deuterium+Enrichment+on+Bacterial+Growth.">Effects of Low-Level Deuterium Enrichment on Bacterial Growth.</a> Plos One. 9 (7), e102071 (2014).</li><li>Okamoto, A., Hashimoto, K., Nealson, K. H., Nakamura, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rate+enhancement+of+bacterial+extracellular+electron+transport+involves+bound+flavin+semiquinones.">Rate enhancement of bacterial extracellular electron transport involves bound flavin semiquinones.</a> Proc. Natl. Acad. Sci. U. S. A. 110 (19), 7856-7861 (2013).</li><li>Edwards, M. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Redox+Linked+Flavin+Sites+in+Extracellular+Decaheme+Proteins+Involved+in+Microbe-Mineral+Electron+Transfer.">Redox Linked Flavin Sites in Extracellular Decaheme Proteins Involved in Microbe-Mineral Electron Transfer.</a> Sci. Rep. 5, 11677 (2015).</li><li>Saito, J., Hashimoto, K., Okamoto, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Flavin+as+an+Indicator+of+the+Rate-Limiting+Factor+for+Microbial+Current+Production+in+Shewanella+oneidensis+MR-1.">Flavin as an Indicator of the Rate-Limiting Factor for Microbial Current Production in Shewanella oneidensis MR-1.</a> Electrochim. Acta. 216, 261-265 (2016).</li><li>Guo, J. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reduction+of+Cr(VI)+by+Escherichia+coli+BL21+in+the+presence+of+redox+mediators.">Reduction of Cr(VI) by Escherichia coli BL21 in the presence of redox mediators.</a> Bioresource Technol. 123, 713-716 (2012).</li><li>Nealson, K., Saffarini, D., Moser, D., Smith, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Spectrophotometric+Method+for+Monitoring+Tactic+Responses+of+Bacteria+under+Anaerobic+Conditions.">A Spectrophotometric Method for Monitoring Tactic Responses of Bacteria under Anaerobic Conditions.</a> J Microbiol. Meth. 20 (3), 211-218 (1994).</li><li>Myers, C. R., Myers, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+surface+exposure+of+the+outer+membrane+cytochromes+of+Shewanella+oneidensis+MR-1.">Cell surface exposure of the outer membrane cytochromes of Shewanella oneidensis MR-1.</a> Lett. Appl. Microbiol. 37 (3), 254-258 (2003).</li><li>Lower, B. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antibody+Recognition+Force+Microscopy+Shows+that+Outer+Membrane+Cytochromes+OmcA+and+MtrC+Are+Expressed+on+the+Exterior+Surface+of+Shewanella+oneidensis+MR-1.">Antibody Recognition Force Microscopy Shows that Outer Membrane Cytochromes OmcA and MtrC Are Expressed on the Exterior Surface of Shewanella oneidensis MR-1.</a> Appl. Environ. Microbiol. 75 (9), 2931-2935 (2009).</li><li>Chen, X. X., Ferrigno, R., Yang, J., Whitesides, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Redox+properties+of+cytochrome+c+adsorbed+on+self-assembled+monolayers:+A+probe+for+protein+conformation+and+orientation.">Redox properties of cytochrome c adsorbed on self-assembled monolayers: A probe for protein conformation and orientation.</a> Langmuir. 18 (18), 7009-7015 (2002).</li><li>McMillan, D. G. G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+impact+of+enzyme+orientation+and+electrode+topology+on+the+catalytic+activity+of+adsorbed+redox+enzymes.">The impact of enzyme orientation and electrode topology on the catalytic activity of adsorbed redox enzymes.</a> Electrochim. Acta. 110, 79-85 (2013).</li><li>Dinh, H. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Iron+corrosion+by+novel+anaerobic+microorganisms.">Iron corrosion by novel anaerobic microorganisms.</a> Nature. 427 (6977), 829-832 (2004).</li><li>McGlynn, S. E., Chadwick, G. L., Kempes, C. P., Orphan, V. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Single+cell+activity+reveals+direct+electron+transfer+in+methanotrophic+consortia.">Single cell activity reveals direct electron transfer in methanotrophic consortia.</a> Nature. 526 (7574), 531-535 (2015).</li><li>Okamoto, A., Nakamura, R., Nealson, K. H., Hashimoto, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bound+Flavin+Model+Suggests+Similar+Electron-Transfer+Mechanisms+in+Shewanella+and+Geobacter.">Bound Flavin Model Suggests Similar Electron-Transfer Mechanisms in Shewanella and Geobacter.</a> Chemelectrochem. 1 (11), 1808-1812 (2014).</li><li>Okamoto, A., Hashimoto, K., Nealson, K. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Flavin+Redox+Bifurcation+as+a+Mechanism+for+Controlling+the+Direction+of+Electron+Flow+during+Extracellular+Electron+Transfer.">Flavin Redox Bifurcation as a Mechanism for Controlling the Direction of Electron Flow during Extracellular Electron Transfer.</a> Angew. Chem. Int. Ed. 53 (41), 10988-10991 (2014).</li><li>Tokunou, Y., Hashimoto, K., Okamoto, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acceleration+of+Extracellular+Electron+Transfer+by+Alternative+Redox-Active+Molecules+to+Riboflavin+for+Outer-Membrane+Cytochrome+c+of+Shewanella+oneidensis+MR-1.">Acceleration of Extracellular Electron Transfer by Alternative Redox-Active Molecules to Riboflavin for Outer-Membrane Cytochrome c of Shewanella oneidensis MR-1.</a> J Phys. Chem. C. 120 (29), 16168-16173 (2016).</li><li>Rowe, A. 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Vår hela-cell elektrokemisk analys har flera tekniska fördelar jämfört med protein elektrokemi. Medan protein rening kräver flera steg tidsödande förfaranden, tar vår hela-cell metoden en dag i självorganiserade biofilm bildning efter cellodling. För att uppnå en stabil interaktion mellan OM c- Cyts och elektroden, vi behöver bara sterilisering och rengöring av elektrod ytan; det inte kräver ändringar av elektrod för organisera orienteringen av proteiner4, t.ex., ...

Elektrokemiska tekniker att direkt karakterisera en redox protein i en intakt bakteriecell har nyligen dykt upp sedan upptäckten av metall-minska mikrobiell stammar, såsom S. oneidensis MR-1 eller Geobacter sulfurreducens PCA, som har yttre membran typ c cytokrom komplex (OM c-Cyts) utsätts för cell exteriör1,2,3,4,5. OM c- Cyts medla elektrontransport från luftvägarna kedjan till fasta substrat ligger extracellularly. Denna transport kallas extracellulära elektrontransport (EET)1,6 och är en kritisk process för framväxande bioteknik, till exempel mikrobiella bränsleceller6. Därför, för att förstå den underliggande EET-kineticsen och mekanismer, och dess koppling till mikrobiell fysiologi, OM c -Cyts har undersökts med hjälp av hela-cell elektrokemi4,7, kombinerat med mikroskopi 8 , 9, spektroskopi10,11och molekylär biologi2,4. Däremot har metoder för att undersöka effekterna av EET-associerade katjontransport, t.ex., protoner, på EET kinetics i levande celler knappt fastställts, trots proton transport över bakteriell membran har en avgörande roll i signalering, homeostas och energi produktion12,13,14. I denna studie, beskriver vi en teknik för att undersöka effekterna av proton transport på EET kinetics i S. oneidensis MR-1 cellen använder hela-cell elektrokemiska mätningar, som kräver identifiering av det hastighetsbegränsande steget i mikrobiell nuvarande produktion15.
Ett direkt sätt att utvärdera bidraget av proton transporter på den associerade EET är effekten kinetiska isotopen deuterium (KIE). KIE är observerbar som förändringen i elektron överföring kinetik vid byte av protoner med deuterium joner, som representerar proton transporternas inverkan på elektron överföring kinetik16. Teorin om KIE själv har väl etablerad med elektrokemiska mätningar med renade enzymer17. Emellertid eftersom nuvarande produktion i S. oneidensis MR-1 resultat från flera olika och varierande processer18, identifiera inte ett helt enkelt EET som hastighetsbegränsning processen. För att observera KIE på proton processer tillsammans med EET, måste vi bekräfta att den mikrobiella aktuella produktionen begränsas av elektrontransport via OM c- Cyts till elektroden. För detta ändamål ersatte vi supernatanten med färska medium som innehåller en hög koncentration av laktat som en elektron donator vid optimalt pH och temperatur före KIE mätning; Detta utbyte serveras två roller: (1) den förbättrade graden av den uppströms metaboliska processer jämfört med EET och (2) utelämnas simning cellerna i supernatanten frigörs från den enskiktslager biofilmen av S. oneidensis MR-1 på den arbetande elektrod ( indium tin-dopade oxid (ITO) elektrod). Presenterade detaljerade protokollet är avsett att hjälpa nya utövare upprätthålla och bekräfta att EET processen är det kurs-bestämma steget.

<table>
	<tbody>
		<tr>
			<td>Glass cylinder</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>Custom-made, used as the electrochemical reactor</td>
		</tr>
		<tr>
			<td>PTFE cover and base</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>Custom-made, used as a cover and a foundation of the electrochemical reactor</td>
		</tr>
		<tr>
			<td>Buthyl rubber</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>Custom-made, inserted between each component of electrochemical reactor</td>
		</tr>
		<tr>
			<td>Septa</td>
			<td>GL Science</td>
			<td>3007-16101</td>
			<td>Used as an injection port of electrochemical reactor</td>
		</tr>
		<tr>
			<td>Indium tin-doped oxide (ITO) electrode</td>
			<td>GEOMATEC</td>
			<td>No.0001</td>
			<td>Used as a working electrode, 5&#937;/sq</td>
		</tr>
		<tr>
			<td>Ag/AgCl KCl saturated electrode</td>
			<td>HOKUTO DENKO</td>
			<td>HX-R5</td>
			<td>Used as a reference electrode, &#934;0.30mm</td>
		</tr>
		<tr>
			<td>Platinum wire</td>
			<td>The Nilaco Cooporation</td>
			<td>PT-351325</td>
			<td>Used as a counter electrode</td>
		</tr>
		<tr>
			<td>Luria-Bertani (LB) Broth, Miller</td>
			<td>Becton, Dichkinson and Company</td>
			<td>244620</td>
			<td>Medium for precultivation of S. oneidensis MR-1</td>
		</tr>
		<tr>
			<td>Bacto agar</td>
			<td>Becton, Dichkinson and Company</td>
			<td>214010</td>
			<td></td>
		</tr>
		<tr>
			<td>Anthraquinone-1-sulfonate (&#945;-AQS)</td>
			<td>TCI</td>
			<td>A1428</td>
			<td></td>
		</tr>
		<tr>
			<td>Flavin mononucleotide (FMN)</td>
			<td>Wako</td>
			<td>184-00831</td>
			<td></td>
		</tr>
		<tr>
			<td>NaHCO3</td>
			<td>Wako</td>
			<td>191-01305</td>
			<td>Used for defined medium (DM)</td>
		</tr>
		<tr>
			<td>CaCl2 &#183; 2H2O</td>
			<td>Wako</td>
			<td>031-00435</td>
			<td>Used for DM</td>
		</tr>
		<tr>
			<td>NH4Cl</td>
			<td>Wako</td>
			<td>011-03015</td>
			<td>Used for DM</td>
		</tr>
		<tr>
			<td>MgCl2 &#183; 6H2O</td>
			<td>Wako</td>
			<td>135-00165</td>
			<td>Used for DM</td>
		</tr>
		<tr>
			<td>NaCl</td>
			<td>Wako</td>
			<td>191-01665</td>
			<td>Used for DM</td>
		</tr>
		<tr>
			<td>2-[4-(2-hydroxyethyl)-1-piperazinyl] ethanesulfonic acid (HEPES)</td>
			<td>DOJINDO</td>
			<td>346-08235</td>
			<td>Used for DM</td>
		</tr>
		<tr>
			<td>Sodium Lactate Solution</td>
			<td>Wako</td>
			<td>195-02305</td>
			<td></td>
		</tr>
		<tr>
			<td>Bacto Yeast Extract</td>
			<td>Becton, Dichkinson and Company</td>
			<td>212750</td>
			<td></td>
		</tr>
		<tr>
			<td>Deuterium oxide (D, 99.9%)</td>
			<td>Cambridge Isotope Laboratories, Inc.</td>
			<td>DLM-4-PK</td>
			<td>Additive for kinetic isotope effect experiments</td>
		</tr>
		<tr>
			<td>Incubator</td>
			<td>TOKYO RIKAKIKAI CO. LTD.</td>
			<td>LTI-601SD</td>
			<td>Used for precultivation</td>
		</tr>
		<tr>
			<td>Shaker</td>
			<td>TAITEC</td>
			<td>NR-3</td>
			<td>Used for precultivation</td>
		</tr>
		<tr>
			<td>Autoclave machine</td>
			<td>TOMY SEIKO CO. LTD.</td>
			<td>LSX-500</td>
			<td>Used for sterilization of the electrochemical reactor and the medium</td>
		</tr>
		<tr>
			<td>Clean bench</td>
			<td>SANYO</td>
			<td>MCV-91BNF</td>
			<td>Used to prevent the contamination of the electrochemical reactor and the medium with other microbes</td>
		</tr>
		<tr>
			<td>Centrifuge separator</td>
			<td>Eppendorf</td>
			<td>5430R</td>
			<td>Rotational speed upto 6000&#215;g is required</td>
		</tr>
		<tr>
			<td>Nitrogen gas generator</td>
			<td>Puequ CO. LTD.</td>
			<td>PNTN-2</td>
			<td>Nitrogen gas cylinder can also be used instead of gas generator</td>
		</tr>
		<tr>
			<td>UV-vis spectrometer</td>
			<td>SHIMADZU</td>
			<td>UV-1800</td>
			<td>Used for optimization of cell density</td>
		</tr>
		<tr>
			<td>Potentiostat</td>
			<td>BioLogic</td>
			<td>VMP3</td>
			<td>Used for biofilm formation and kinetic isotope effect experiments</td>
		</tr>
		<tr>
			<td>Thermal water circulator</td>
			<td>AS ONE</td>
			<td>TR-1A</td>
			<td>Used for maintanance of temperature of electrochemcial reactor</td>
		</tr>
		<tr>
			<td>Faraday cage</td>
			<td>HOKUTO DENKO</td>
			<td>HS-201S</td>
			<td>Used for electrochemical experiments</td>
		</tr>
	</tbody>
</table>

electrochemical detection of deuterium kinetic isotope effect on extracellular electron transport in shewanella oneidensis mr-1

Direkt elektrokemisk detektion av c-Skriv cytokrom komplex inbäddad i den bakteriella yttre membranen (yttre membran c-Skriv cytokrom komplex; OM c- Cyts) har nyligen dykt upp som en roman hela-cell analysmetod att karakterisera den bakteriella elektrontransport från luftvägarna kedjan på cell utsidan, kallad den extracellulära elektrontransport (EET). Medan de utbildningsavsnittet och kinetik av elektronflödet under EET reaktionen har undersökts, har en hela-cell elektrokemisk metod att undersöka effekterna av katjontransport som är associerad med EET ännu inte fastställts. I den aktuella studien, ett exempel på en biokemisk teknik för att undersöka deuterium kinetiska isotopen effekten (KIE) på EET genom OM c- Cyts använder en modell mikrob, Shewanella oneidensis MR-1, är beskriven. KIE på EET processen kan erhållas om EET genom OM c- Cyts fungerar som det hastighetsbegränsande steget i den mikrobiella aktuella produktionen. I detta syfte före tillsats av D2O, ersattes supernatanten med färska media som innehåller en tillräcklig mängd av elektron donator att stödja frekvensen av uppströms metaboliska reaktioner, och ta bort plankton cellerna från en enhetlig enskiktslager biofilm på arbetselektroden. Alternativa metoder för att bekräfta begränsa steg i mikrobiell nuvarande produktion som EET genom OM c- Cyts beskrivs också. Vår teknik med en hela-cell elektrokemisk analys för att undersöka proton transport kinetik kan tillämpas på andra elektroaktiva mikrobiell stammar.

Efter 25 h av potentiella ansökan vid 0,4 V (kontra hon) bildades en enskiktslager biofilm på arbetselektroden ITO glas, vilket bekräftades tidigare av antingen en scanning electron microscopy eller en konfokalmikroskopi4. Representativa tidsförloppet för aktuell produktion från S. oneidensis herr-1 under bildandet av en enskiktslager biofilm visas i figur 2. Även om nuvarande förändrar i varje mätning, uppvisar den ...

Watch this Scientific Journal Video about Electrochemical Detection of Deuterium Kinetic Isotope Effect on Extracellular Electron Transport in Shewanella oneidensis MR-1 at JoVE.com

Electrochemical Detection of Deuterium Kinetic Isotope Effect on Extracellular Electron Transport in Shewanella oneidensis MR-1

Elektrokemisk detektion av Deuterium Kinetic isotopen effekt på extracellulära elektrontransport i Shewanella oneidensis MR-1

Här presenterar vi ett protokoll av hela-cell elektrokemiska experiment för att studera bidrag proton transport till graden av extracellulära elektrontransport via de yttre-membran subfamiljen komplex i Shewanella oneidensis MR-1.

Direct electrochemical detection of c-type cytochrome complexes embedded in the bacterial outer membrane (outer membrane c-type cytochrome complexes; OM ...

Research

JoVE Journal

Biochemistry

Electrochemical Detection of Deuterium Kinetic Isotope Effect on Extracellular Electron Transport in Shewanella oneidensis MR-1

Reconstitution of Nucleosomes with Differentially Isotope-labeled Sister Histones

Beredning av Nukleosomer med differentiellt isotopmärkt Sister Histoner

Asymmetrically modified nucleosomes contain the two copies of a histone (sister histones) decorated with distinct sets of Post-translational Modifications ...

Time-resolved ElectroSpray Ionization Hydrogen-deuterium Exchange Mass Spectrometry for Studying Protein Structure and Dynamics

Tidsupplöst elektrosprayjonisering Väte-deuterium Exchange-masspektrometri för att studera Protein Structure and Dynamics

Intrinsically disordered proteins (IDPs) have long been a challenge to structural biologists due to their lack of stable secondary structure elements. ...

Analyzing Supercomplexes of the Mitochondrial Electron Transport Chain with Native Electrophoresis, In-gel Assays, and Electroelution

Analysera superkomplex av den mitokondriella elektrontransportkedjan med naturliga elektrofores, in-gel-analyser och elektrolys

The mitochondrial electron transport chain (ETC) transduces the energy derived from the breakdown of various fuels into the bioenergetic currency of the ...

A Simple Method for High Throughput Chemical Screening in Caenorhabditis Elegans

A Simple Method for High Throughput Chemical Screening in Caenorhabditis Elegans

En enkel metod för hög genomströmning kemisk Screening i Caenorhabditis Elegans

Caenorhabditis elegans is a useful organism for testing chemical effects on physiology. Whole organism small molecule screens offer significant advantages ...

Measuring Trans-Plasma Membrane Electron Transport by C2C12 Myotubes

Mäta Trans-Plasma membran elektrontransport av C2C12 Myotubes

Trans-plasma membrane electron transport (tPMET) plays a role in protection of cells from intracellular reductive stress as well as protection from damage ...

Defining Hsp33's Redox-regulated Chaperone Activity and Mapping Conformational Changes on Hsp33 Using Hydrogen-deuterium Exchange Mass Spectrometry

Definiera Hsp33's Redox-reglerade förkläde aktivitet och mappning konfirmerande ändringar på Hsp33 med hjälp av väte-deuterium Exchange masspektrometri

Living organisms regularly need to cope with fluctuating environments during their life cycle, including changes in temperature, pH, the accumulation of ...

Sampling and Pretreatment of Tooth Enamel Carbonate for Stable Carbon and Oxygen Isotope Analysis

Provtagning och förbehandling av tandemaljen karbonat för stabil kol och syre isotoper analys

Stable carbon and oxygen isotope analysis of human and animal tooth enamel carbonate has been applied in paleodietary, paleoecological, and ...

Extracellular Protein Microarray Technology for High Throughput Detection of Low Affinity Receptor-Ligand Interactions

Extracellulära Protein Mikroarrayteknik för hög genomströmning upptäckt av låg affinitet Receptor-Ligand interaktioner

Secreted factors, membrane-tethered receptors, and their interacting partners are main regulators of cellular communication and initiation of signaling ...

Analysis of &#946;-Amyloid-induced Abnormalities on Fibrin Clot Structure by Spectroscopy and Scanning Electron Microscopy

Analysis of ÃƒÆ’Ã…Â½Ãƒâ€šÃ‚Â²-Amyloid-induced Abnormalities on Fibrin Clot Structure by Spectroscopy and Scanning Electron Microscopy

Analys av β-Amyloid-inducerad avvikelser på Fibrin Clot struktur av spektroskopi och svepelektronmikroskopi

This article presents methods for generating in vitro fibrin clots and analyzing the effect of beta-amyloid (A&#946;) protein on clot formation and ...

Analysis of Protein Folding, Transport, and Degradation in Living Cells by Radioactive Pulse Chase

Analys av proteinveckning, Transport och nedbrytning i levande celler av radioaktiva puls Chase

Radioactive pulse-chase labeling is a powerful tool for studying the conformational maturation, the transport to their functional cellular location, and ...