Sökning: WFRF:(Barwell J)
> (2010-2014) >
Production, Purific...
Production, Purification and Characterization of Recombinant, Full-Length Human Claudin-1
-
- Bonander, Nicklas, 1968 (författare)
- Chalmers tekniska högskola,Chalmers University of Technology
-
- Jamshad, M. (författare)
- Aston University
-
- Oberthür, D. (författare)
- Center for Free-Electron Laser Science (CFEL),Universität Hamburg,University of Hamburg
-
visa fler...
-
- Clare, M. (författare)
- Aston University
-
- Barwell, J. (författare)
- Aston University
-
- Hu, K. (författare)
- University of Birmingham
-
- Farquhar, M.J. (författare)
- University of Birmingham
-
- Stamataki, Z. (författare)
- University of Birmingham
-
- Harris, H.J. (författare)
- University of Birmingham
-
- Dierks, K. (författare)
- Universität Hamburg,University of Hamburg
-
- Dafforn, T.R. (författare)
- University of Birmingham
-
- Betzel, C. (författare)
- Universität Hamburg,University of Hamburg
-
- McKeating, J.A. (författare)
- University of Birmingham
-
- Bill, R (författare)
- Aston University
-
visa färre...
-
(creator_code:org_t)
- 2013-05-21
- 2013
- Engelska.
-
Ingår i: PLoS ONE. - : Public Library of Science (PLoS). - 1932-6203 .- 1932-6203. ; 8:5, s. Art. no. e64517-
- Relaterad länk:
-
http://publications.... (primary) (free)
-
visa fler...
-
https://journals.plo...
-
https://doi.org/10.1...
-
https://research.cha...
-
visa färre...
Abstract
Ämnesord
Stäng
- The transmembrane domain proteins of the claudin superfamily are the major structural components of cellular tight junctions. One family member, claudin-1, also associates with tetraspanin CD81 as part of a receptor complex that is essential for hepatitis C virus (HCV) infection of the liver. To understand the molecular basis of claudin-1/CD81 association we previously produced and purified milligram quantities of functional, full-length CD81, which binds a soluble form of HCV E2 glycoprotein (sE2). Here we report the production, purification and characterization of claudin-1. Both yeast membrane-bound and detergent-extracted, purified claudin-1 were antigenic and recognized by specific antibodies. Analytical ultracentrifugation demonstrated that extraction with n-octyl-β-d-glucopyranoside yielded monodispersed, dimeric pools of claudin-1 while extraction with profoldin-8 or n-decylphosphocholine yielded a dynamic mixture of claudin-1 oligomers. Neither form bound sE2 in line with literature expectations, while further functional analysis was hampered by the finding that incorporation of claudin-1 into proteoliposomes rendered them intractable to study. Dynamic light scattering demonstrated that claudin-1 oligomers associate with CD81 in vitro in a defined molar ratio of 1:2 and that complex formation was enhanced by the presence of cholesteryl hemisuccinate. Attempts to assay the complex biologically were limited by our finding that claudin-1 affects the properties of proteoliposomes. We conclude that recombinant, correctly-folded, full-length claudin-1 can be produced in yeast membranes, that it can be extracted in different oligomeric forms that do not bind sE2 and that a dynamic preparation can form a specific complex with CD81 in vitro in the absence of any other cellular components. These findings pave the way for the structural characterization of claudin-1 alone and in complex with CD81.
Ämnesord
- NATURVETENSKAP -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)
Publikations- och innehållstyp
- art (ämneskategori)
- ref (ämneskategori)
Hitta via bibliotek
-
PLoS ONE
(Sök värdpublikationen i LIBRIS)
Till lärosätets databas
- Av författaren/redakt...
-
Bonander, Nickla ...
-
Jamshad, M.
-
Oberthür, D.
-
Clare, M.
-
Barwell, J.
-
Hu, K.
-
visa fler...
-
Farquhar, M.J.
-
Stamataki, Z.
-
Harris, H.J.
-
Dierks, K.
-
Dafforn, T.R.
-
Betzel, C.
-
McKeating, J.A.
-
Bill, R
-
visa färre...
- Om ämnet
-
- NATURVETENSKAP
-
NATURVETENSKAP
-
och Biologi
-
och Biokemi och mole ...
- Artiklar i publikationen
-
PLoS ONE
- Av lärosätet
-
Chalmers tekniska högskola