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  4. Reengineering of the human pyruvate dehydrogenase complex: from disintegration to highly active agglomerates
 
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Reengineering of the human pyruvate dehydrogenase complex: from disintegration to highly active agglomerates

Publikationstyp
Journal Article
Date Issued
2016-12-16
Sprache
English
Author(s)
Guo, Jin  
Hezaveh, Samira  
Tatur, Jana  
Zeng, An-Ping  orcid-logo
Jandt, Uwe  
Institut
Bioprozess- und Biosystemtechnik V-1  
TORE-URI
http://hdl.handle.net/11420/3241
Journal
Biochemical journal  
Volume
474
Issue
5
Start Page
865
End Page
875
Citation
Biochemical Journal 5 (474): 865-875 (2017-03-01)
Publisher DOI
10.1042/BCJ20160916
Scopus ID
2-s2.0-85014718328
Publisher
Portland Press
The pyruvate dehydrogenase complex (PDC) plays a central role in cellular metabolism and regulation. As a metabolite-channeling multi-enzyme complex it acts as a complete nanomachine due to its unique geometry and by coupling a cascade of catalytic reactions using 'swinging arms'. Mammalian and specifically human PDC (hPDC) is assembled from multiple copies of E1 and E3 bound to a large E2/E3BP 60-meric core. A less restrictive and smaller catalytic core, which is still active, is highly desired for both fundamental research on channeling mechanisms and also to create a basis for further modification and engineering of new enzyme cascades. Here, we present the first experimental results of the successful disintegration of the E2/E3BP core while retaining its activity. This was achieved by C-terminal α-helixes double truncations (eight residues from E2 and seven residues from E3BP). Disintegration of the hPDC core via double truncations led to the formation of highly active (approximately 70% of wildtype) apparently unordered clusters or agglomerates and inactive non-agglomerated species (hexamer/ trimer). After additional deletion of N-terminal 'swinging arms', the aforementioned C-terminal truncations also caused the formation of agglomerates of minimized E2/E3BP complexes. It is likely that these 'swinging arm' regions are not solely responsible for the formation of the large agglomerates.
DDC Class
570: Biowissenschaften, Biologie
610: Medizin
620: Ingenieurwissenschaften
More Funding Information
Funded by the German Federal Ministry of Education and Research (BMBF, grant [031A128]). The research leading to these results has received funding from the European Community's Seventh Framework Programme (FP7/2007-2013) under BioStruct-X (grant agreement No 283570).
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