The rat UCP1 (uncoupling protein 1) is a mitochondrial inner-membrane carrier involved in energy dissipation and heat production. We expressed UCP1 carrying a His6 epitope at its C-terminus in Saccharomyces cerevisiae mitochondria. The recombinant-tagged UCP1 was purified by immobilized metal-ion affinity chromatography to homogeneity (>95%). This made it suitable for subsequent biophysical characterization. Fluorescence resonance energy transfer experiments showed that n-dodecyl-β-d-maltoside-solubilized UCP1–His6 retained its PN (purine nucleotide)-binding capacity. The far-UV CD spectrum of the functional protein clearly indicated the predominance of α-helices in the UCP1 secondary structure. The UCP1 secondary structure exhibited an α-helical degree of approx. 68%, which is at least 25% higher than the previously reported estimations based on computational predictions. Moreover, the helical content remained unchanged in free and PN-loaded UCP1. A homology model of the first repeat of UCP1, built on the basis of X-ray-solved close parent, the ADP/ATP carrier, strengthened the CD experimental results. Our experimental and computational results indicate that (i) α-helices are the major component of UCP1 secondary structure; (ii) PN-binding mechanism does not involve significant secondary-structure rearrangement; and (iii) UCP1 shares similar secondary-structure characteristics with the ADP/ATP carrier, at least for the first repeat.
Skip Nav Destination
Follow us on Twitter @Biochem_Journal
Article navigation
Research Article|
May 15 2004
Secondary-structure characterization by far-UV CD of highly purified uncoupling protein 1 expressed in yeast Available to Purchase
Pierre DOUETTE;
Pierre DOUETTE
*Laboratory of Bioenergetics, Centre for Oxygen Research and Development, Institute of Chemistry B6, University of Liege, Sart Tilman, B-4000 Liege, Belgium
Search for other works by this author on:
Rachel NAVET;
Rachel NAVET
*Laboratory of Bioenergetics, Centre for Oxygen Research and Development, Institute of Chemistry B6, University of Liege, Sart Tilman, B-4000 Liege, Belgium
Search for other works by this author on:
Fabrice BOUILLENNE;
Fabrice BOUILLENNE
†Centre for Protein Engineering, Institute of Chemistry B6, University of Liege, Sart Tilman, B-4000 Liege, Belgium
Search for other works by this author on:
Alain BRANS;
Alain BRANS
†Centre for Protein Engineering, Institute of Chemistry B6, University of Liege, Sart Tilman, B-4000 Liege, Belgium
Search for other works by this author on:
Claudine SLUSE-GOFFART;
Claudine SLUSE-GOFFART
*Laboratory of Bioenergetics, Centre for Oxygen Research and Development, Institute of Chemistry B6, University of Liege, Sart Tilman, B-4000 Liege, Belgium
Search for other works by this author on:
André MATAGNE;
André MATAGNE
†Centre for Protein Engineering, Institute of Chemistry B6, University of Liege, Sart Tilman, B-4000 Liege, Belgium
Search for other works by this author on:
Francis E. SLUSE
Francis E. SLUSE
1
*Laboratory of Bioenergetics, Centre for Oxygen Research and Development, Institute of Chemistry B6, University of Liege, Sart Tilman, B-4000 Liege, Belgium
1To whom correspondence should be addressed (e-mail [email protected]).
Search for other works by this author on:
Publisher: Portland Press Ltd
Received:
December 19 2003
Revision Received:
February 05 2004
Accepted:
February 06 2004
Accepted Manuscript online:
February 06 2004
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London ©2004
2004
Biochem J (2004) 380 (1): 139–145.
Article history
Received:
December 19 2003
Revision Received:
February 05 2004
Accepted:
February 06 2004
Accepted Manuscript online:
February 06 2004
Citation
Pierre DOUETTE, Rachel NAVET, Fabrice BOUILLENNE, Alain BRANS, Claudine SLUSE-GOFFART, André MATAGNE, Francis E. SLUSE; Secondary-structure characterization by far-UV CD of highly purified uncoupling protein 1 expressed in yeast. Biochem J 15 May 2004; 380 (1): 139–145. doi: https://doi.org/10.1042/bj20031957
Download citation file:
Sign in
Don't already have an account? Register
Sign in to your personal account
You could not be signed in. Please check your email address / username and password and try again.
Could not validate captcha. Please try again.
Biochemical Society Member Sign in
Sign InSign in via your Institution
Sign in via your InstitutionGet Access To This Article
Follow us on Twitter @Biochem_Journal
Open Access for all
We offer compliant routes for all authors from 2025. With library support, there will be no author nor reader charges in 5 journals. Check here |
![]() View past webinars > |