Sökning: onr:"swepub:oai:DiVA.org:uu-232888" > Expression, purific...
Fältnamn | Indikatorer | Metadata |
---|---|---|
000 | 04481naa a2200721 4500 | |
001 | oai:DiVA.org:uu-232888 | |
003 | SwePub | |
008 | 140926s2014 | |||||||||||000 ||eng| | |
024 | 7 | a https://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-2328882 URI |
024 | 7 | a https://doi.org/10.1107/S20522525140149002 DOI |
040 | a (SwePub)uu | |
041 | a engb eng | |
042 | 9 SwePub | |
072 | 7 | a ref2 swepub-contenttype |
072 | 7 | a art2 swepub-publicationtype |
100 | 1 | a Lee, Ho-Hsien4 aut |
245 | 1 0 | a Expression, purification and crystallization of CTB-MPR, a candidate mucosal vaccine component against HIV-1 |
264 | 1 | c 2014 |
338 | a print2 rdacarrier | |
520 | a CTB-MPR is a fusion protein between the B subunit of cholera toxin (CTB) andthe membrane-proximal region of gp41 (MPR), the transmembrane envelopeprotein ofHuman immunodeficiency virus 1(HIV-1), and has previously beenshown to induce the production of anti-HIV-1 antibodies with antiviralfunctions. To further improve the design of this candidate vaccine, X-raycrystallography experiments were performed to obtain structural informationabout this fusion protein. Several variants of CTB-MPR were designed,constructed and recombinantly expressed inEscherichia coli. The first variantcontained a flexible GPGP linker between CTB and MPR, and yielded crystalsthat diffracted to a resolution of 2.3 A ̊, but only the CTB region was detectedin the electron-density map. A second variant, in which the CTB was directlyattached to MPR, was shown to destabilize pentamer formation. A thirdconstruct containing a polyalanine linker between CTB and MPR proved tostabilize the pentameric form of the protein during purification. The purificationprocedure was shown to produce a homogeneously pure and monodispersesample for crystallization. Initial crystallization experiments led to pseudo-crystals which were ordered in only two dimensions and were disordered inthe third dimension. Nanocrystals obtained using the same precipitant showedpromising X-ray diffraction to 5 A ̊resolution in femtosecond nanocrystallo-graphy experiments at the Linac Coherent Light Source at the SLAC NationalAccelerator Laboratory. The results demonstrate the utility of femtosecondX-ray crystallography to enable structural analysis based on nano/microcrystalsof a protein for which no macroscopic crystals ordered in three dimensions havebeen observed before. | |
650 | 7 | a NATURVETENSKAPx Biologix Biokemi och molekylärbiologi0 (SwePub)106022 hsv//swe |
650 | 7 | a NATURAL SCIENCESx Biological Sciencesx Biochemistry and Molecular Biology0 (SwePub)106022 hsv//eng |
700 | 1 | a Cherni, Irene4 aut |
700 | 1 | a Yu, HongQi4 aut |
700 | 1 | a Fromme, Raimund4 aut |
700 | 1 | a Doran, Jeffrey D.4 aut |
700 | 1 | a Grotjohann, Ingo4 aut |
700 | 1 | a Mittman, Michele4 aut |
700 | 1 | a Basu, Shibom4 aut |
700 | 1 | a Deb, Arpan4 aut |
700 | 1 | a Dörner, Katerina4 aut |
700 | 1 | a Aquila, Andrew4 aut |
700 | 1 | a Barty, Anton4 aut |
700 | 1 | a Boutet, Sébastien4 aut |
700 | 1 | a Chapman, Henry N.4 aut |
700 | 1 | a Doak, R. Bruce4 aut |
700 | 1 | a Hunter, Mark S.4 aut |
700 | 1 | a James, Daniel4 aut |
700 | 1 | a Kirian, Richard A.4 aut |
700 | 1 | a Kupitz, Christopher4 aut |
700 | 1 | a Lawrence, Robert M.4 aut |
700 | 1 | a Liu, Haiguang4 aut |
700 | 1 | a Nass, Karol4 aut |
700 | 1 | a Schlichting, Ilme4 aut |
700 | 1 | a Schmidt, Kevin E.4 aut |
700 | 1 | a Seibert, M. Marvinu Linac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, CA 94025, USA,4 aut0 (Swepub:uu)masei255 |
700 | 1 | a Shoeman, Robert L.4 aut |
700 | 1 | a Spence, John C. H.4 aut |
700 | 1 | a Stellato, Francesco4 aut |
700 | 1 | a Weierstall, Uwe4 aut |
700 | 1 | a Williams, Garth J.4 aut |
700 | 1 | a Yoon, Chunhong4 aut |
700 | 1 | a Wang, Dingjie4 aut |
700 | 1 | a Zatsepin, Nadia A.4 aut |
700 | 1 | a Hogue, Brenda G.4 aut |
700 | 1 | a Matoba, Nobuyuki4 aut |
700 | 1 | a Fromme, Petra4 aut |
700 | 1 | a Mor, Tsafrir S.4 aut |
710 | 2 | a Linac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, CA 94025, USA,4 org |
773 | 0 | t IUCrJg 1:5, s. 305-317q 1:5<305-317x 2052-2525 |
856 | 4 | u http://dx.doi.org/10.1107/S2052252514014900 |
856 | 4 8 | u https://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-232888 |
856 | 4 8 | u https://doi.org/10.1107/S2052252514014900 |
Kungliga biblioteket hanterar dina personuppgifter i enlighet med EU:s dataskyddsförordning (2018), GDPR. Läs mer om hur det funkar här.
Så här hanterar KB dina uppgifter vid användning av denna tjänst.