Name: small and wide angle x-ray scattering studies of biological macromolecules in solution
Uploaded: 2013-01-08T12:00:00
Description: Watch this Scientific Journal Video about Small and Wide Angle X-Ray Scattering Studies of Biological Macromolecules in Solution at JoVE.com

Vi vil gerne takke Dr. Manfred Kriechbaum af Hecus Rtg og Institut for Biofysik og Nanosystems Forskning ved den østrigske Academy of Sciences i Graz, Østrig. LL og XW blev delvist understøttet af US Department of Energy, under DMU-C Award nr. DE-FG07-07ID14889 og US Nuclear Regulatory Commission, under Award No NRC-38-08-950. Den SWAXS instrumentet også delvist understøttet af US Department of Energy, under Award nr. DE-NE0000325.

1. Forberedelse af prøven <ol><li> Anvende en kanyle til at fjerne noget af prøven fra prøvebeholderen. * </li><li> Brug nålen til at fylde kapillarrøret (maksimal diameter på 2,2 mm) med prøven. Kapillarrøret skal fyldes mellem 2 og 3 cm fra bunden. </li><li> Luk kapillarrøret ved smeltende voks på sin spids. </li><li> Skru vakuum prøveholderen fra systemet. </li><li> Tag kapillarrøret af den fusionerede ende (voks ende) og sæt un-kondenserede ende i prøveholderen. </li><li> Sæt holderen tilbage i syst...

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	<li>Bernad&#243;, P., Blackledge, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Structural+biology:+Proteins+in+dynamic+equilibrium.">Structural biology: Proteins in dynamic equilibrium.</a> Nature. 468, 1046-1048 (2010).</li><li>Zhang, F., Skoda, M. W. A., Jacobs, R. M. J., Martin, R. A., Martin, C. M., Schreiber, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protein+Interactions+Studied+by+SAXS:+Effect+of+Ionic+Strength+and+Protein+Concentration+for+BSA+in+Aqueous+Solutions.">Protein Interactions Studied by SAXS: Effect of Ionic Strength and Protein Concentration for BSA in Aqueous Solutions.</a> J. Phys. Chem. B. 111, 251-259 (2007).</li><li>Maranas, J. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+environment+on+local+dynamics+of+macromolecules.">The effect of environment on local dynamics of macromolecules.</a> Current Opinion in Colloid &amp; Interface Science. 12, 29-42 (2007).</li><li>Stribeck, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> X-Ray Scattering of Soft Matter. , (2007).</li><li>Mertens, H. D., Svergun, D. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Structural+characterization+of+proteins+and+complexes+using+small-angle+X-ray+solution+scattering.">Structural characterization of proteins and complexes using small-angle X-ray solution scattering.</a> J. Struct. Biol. 172 (1), 128 (2010).</li><li>Svergun, D. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Small-angle+X-ray+and+neutron+scattering+as+a+tool+for+structural+systems+biology.">Small-angle X-ray and neutron scattering as a tool for structural systems biology.</a> Biol. Chem. 391 (7), 737 (2010).</li><li>Putnam, C. D., Hammel, M., Hura, G. L., Tainer, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=X-ray+solution+scattering+(SAXS)+combined+with+crystallography+and+computation:+defining+accurate+macromolecular+structures,+conformations+and+assemblies+in+solution.">X-ray solution scattering (SAXS) combined with crystallography and computation: defining accurate macromolecular structures, conformations and assemblies in solution.</a> Quat. Rev. Biophys. 40, 191-285 (2007).</li><li>Bonini, M., Fratini, E., Baglioni, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=SAXS+study+of+chain-like+structures+formed+by+magnetic+nanoparticles.">SAXS study of chain-like structures formed by magnetic nanoparticles.</a> Materials Science &amp; Engineering C-Biomimetic and Supramolecular Systems. 27 (5-8), 1377-1381 (2007).</li><li>Falletta, E., Ridi, F., Fratini, E., Vannucci, C., Canton, P., Bianchi, S., Castelvetro, V., Baglioni, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+tri-block+copolymer+templated+synthesis+of+gold+nanostructures.">A tri-block copolymer templated synthesis of gold nanostructures.</a> Journal of Colloid and Interface Science. 357 (1), 88-94 (2011).</li><li>Glatter, O., Scherf, G., Schillen, K., Brown, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+a+Poly(ethylene+oxide)+Poly(propylene+oxide)+Triblock+Copolymer+(EO(27)-PO39-EO(27))+in+Aqueous-Solution.">Characterization of a Poly(ethylene oxide) Poly(propylene oxide) Triblock Copolymer (EO(27)-PO39-EO(27)) in Aqueous-Solution.</a> Macromolecules. 27 (21), 6046-6054 (1994).</li><li>Mittelbach, R., Glatter, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+structure+analysis+of+small-angle+scattering+data+from+polydisperse+colloidal+particles.">Direct structure analysis of small-angle scattering data from polydisperse colloidal particles.</a> Journal of Applied Crystallography. 31, 600-608 (1998).</li></ol>

Den sammenlignende fremgangsmåde af SWAXS systemet tillader talrige variabler, der skal bestemmes fra eksperimentel analyse. De parametre, der opnås fra analysen kan anvendes til forskellige formål i henhold til prøven og forsøgsopstillingen. SAXS indeholder oplysninger om nano-skala størrelse og form af objektet, hvorimod WAXS fokuserer på det atomare og mikro-skala struktur (f.eks molekylær gitter, enhedscelle dimension symmetri). Mere specifikt for partikler i fortyndede opløsninger, kan SAXS studer...

<table border="1"><tbody><tr><td> Navn af produkterne </td><td> Firma </td></tr><tr><td> Den system3 små og Wide-Angle X-Ray Scattering (SWAXS) Kamera </td><td> Hecus Rtg og IBN, Graz, Østrig </td></tr><tr><td> Gnom </td><td> ATSAS 2,3 pakke ved D. Svergun EMBL-Hamburg </td></tr></tbody></table>

small and wide angle x-ray scattering studies of biological macromolecules in solution

I dette papir, er lille og Wide Angle X-ray Scattering (SWAXS) analyse af makromolekyler demonstreret gennem eksperimenter. SWAXS er en teknik, hvor røntgenstråler elastisk spredt af en inhomogen stikprøve i nm-området ved små vinkler (typisk 0,1 - 5 °) og brede vinkler (typisk&gt; 5 °). Denne teknik giver oplysninger om form, størrelse og distribution af makromolekyler, karakteristiske afstande delvist bestilt materialer, porestørrelser og overflade-til-volumen-forhold. Small Angle X-ray scattering (SAXS) kan levere strukturel information af makromolekyler mellem 1 og 200 nm, mens Vidvinkel røntgenspredning (WAXS) kan løse endnu mindre Bragg afstand mellem prøver mellem 0,33 nm og 0,49 nm baseret på specifik systemopsætning og detektor. Afstanden bestemmes ud fra Braggs lov og er afhængig af bølgelængden og indfaldsvinklen. I et SWAXS eksperiment, kan materialerne være fasteeller flydende og kan indeholde faste, flydende eller gasformige domæner (såkaldte partikler) af det samme eller et andet materiale i enhver kombination. SWAXS ansøgninger er meget bred og omfatter kolloider af alle typer: metaller, kompositmaterialer, cement, olie, polymerer, plast, proteiner, fødevarer og farmaceutiske produkter. For faste prøver er tykkelsen begrænset til cirka 5 mm. Brug af en lab-baserede SWAXS instrument er nærmere beskrevet i dette dokument. Med den tilgængelige software (f.eks Gnom-ATSAS 2,3 pakke ved D. Svergun EMBL-Hamburg og EasySWAXS software) for SWAXS system, kan et eksperiment udføres for at bestemme visse parametre af interesse for den givne prøve. Et eksempel på et biologisk makromolekyle eksperiment er analysen af ​​2 vægt-% lysozym i en vandbaseret vandig buffer, som kan vælges og fremstilles ved adskillige metoder. Fremstillingen af ​​prøven følger nedenstående retningslinjer i udarbejdelsen af ​​Sample sektion. Gennem SWAXS eksperimenter,vigtige strukturelle parametre for lysozym, f.eks gyrationsradius, analyseres.

SAXS og WAXS helt kan tilvejebringe strukturel information af prøven ved hjælp af følgende parametre: gyrationsradius, partikelstørrelse og form, opløsning struktur faktor, specifik indre overflade og porestørrelse, lattice type og dimension, og elektrontæthed 1. SAXS og WAXS kan også anvendes til at studere protein-dynamik 2. Den strukturelle information af SWAXS eksperimenter opnås ved at sammenligne den eksperimentelt påvist spektre, og de beregningsmæssig...

Watch this Scientific Journal Video about Small and Wide Angle X-Ray Scattering Studies of Biological Macromolecules in Solution at JoVE.com

Small and Wide Angle X-Ray Scattering Studies of Biological Macromolecules in Solution

Demonstrationen af ​​de små og vidvinkel røntgenspredning (SWAXS) procedure er blevet et middel i studiet af biologiske makromolekyler. Gennem brug af instrumentering og procedurer af specifikke vinkler metoder og forberedelse, viser de eksperimentelle data fra SWAXS den atomare og nano-skala karakterisering af makromolekyler.

Små og Wide Angle X-ray scattering Undersøgelser af biologiske makromolekyler i opløsning

In this paper, Small and Wide Angle X-ray Scattering (SWAXS) analysis of macromolecules is demonstrated through experimentation. SWAXS is a technique ...

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