Influenza polymerase heterotrimer expressed in insect cells (MultiBac
technology) as a fully synthetic polyprotein with subunits separated by
TEV sites
TevProtease-Tev-PA-Tev-PB1-Tev-PB2-Tev-CFP
Reich et al. Nature 2014; doi:10.1038/nature14009
/Extended Data Figure 1 | Production and characterization of influenza B//
//polymerase heterotrimer. a, Schematic of the self-cleaving polyprotein//
//construct used to express recombinant influenza B heterotrimeric
polymerase//
//in insect cells. N-terminally it encodes the tobacco etch virus (TEV)
protease//
//that cleaves C-terminal to the amino-acid sequence ENLYFQ (in italics)
and//
//releases N-terminally His-tagged PA, PB1, C-terminally Strep-tagged PB2//
//and cyan fluorescent protein (CFP) for facilitated expression
monitoring.//
//Arrows indicate the N-to-C-terminal direction and the termini of each
mature//
//protein. The histidine and streptavidin tags are underlined./
Darren
On 02/10/2019 10:18, Paula Salgado wrote:
Dear colleagues
We have been working on a protein that is produced as a pro-peptide,
cleaved internally and reassembled into a complex. The interacting
regions are the new C and N-termini at the cleavage site, so no
rearrangement or loss of part of the protein is involved. The
resulting interacting region is quite intricate, with a helix bundle
and beta-sheet composed of members of each partner protein.
I wonder if there are other examples of such processing required for
folding/assembly of a functional complex? I'm not talking of
processing involving removal of termini or internal regions like
signal removal in transported proteins or maturation of preproinsulin,
for example. I'm interested in examples where a propeptide is cleaved,
then the two proteins reassemble in a complex via interactions of the
similarly oriented new termini.
Any suggestions most appreciated.
Thanks a lot!
Paula
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--
**********************************************************************
Dr. Darren J. Hart,
CNRS Research Director, Institut de Biologie Structurale (IBS)
Unité Mixte de Recherche UMR5075 (CEA-CNRS-UGA)
Director, Integrated Structural Biology Grenoble (ISBG)
Unité Mixte de Service UMS3518 (CNRS-UGA-CEA-EMBL)
**********************************************************************
Email: darren.h...@ibs.fr
Tel: +33 4 57 42 85 86
Physical address: IBS/ISBG, 71 avenue des Martyrs, 38000 Grenoble, France
Postal address: IBS/ISBG, 71 avenue des Martyrs, CS 20192, 38042
Grenoble, Cedex 9, France
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