Expression of protein complexes using multiple Escherichia coli protein co-expression systems: A benchmarking study

Busso, Didier; Peleg, Yoav; Heidebrecht, Tatjana; Romier, Christophe; Jacobovitch, Yossi; Dantes, Ada; Salim, Loubna; Troesch, Edouard; Schuetz, A.; Heinemann, Udo; Folkers, Gert E.; Geerlof, Arie; Wilmanns, Matthias; Polewacz, Andrea; Quedenau, Claudia; Büssow, Konrad; Adamson, Rachel; Blagova, E.V.; Walton, Julia; Cartwright, Jared; Bird, Louise E.; Owens, Raymond J.; Berrow, Nicholas S.; Wilson, K.S.; Sussman, Joel L.; Perrakis, Anastassis; Celie, P.H.N.

doi:10.1016/j.jsb.2011.03.004

Cited by 38 publications

(22 citation statements)

References 44 publications

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“…A variety of new technologies for DNA manipulation including ligation independent or restriction free procedures, in-fusion or gateway approaches are now being used in addition to classical restriction-based strategies (see Busso et al, 2011 for examples and test cases). Partner Grenoble has developed a system for combinatorial gene assembly into multigene expression vectors called ACEMBL.…”

Section: Challenges For Sample Preparation: New Methods For Protein Cmentioning

confidence: 99%

Structural insights into transcription complexes

Berger

Blanco

Boelens

et al. 2011

Journal of Structural Biology

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a b s t r a c tControl of transcription allows the regulation of cell activity in response to external stimuli and research in the field has greatly benefited from efforts in structural biology. In this review, based on specific examples from the European SPINE2-COMPLEXES initiative, we illustrate the impact of structural proteomics on our understanding of the molecular basis of gene expression. While most atomic structures were obtained by X-ray crystallography, the impact of solution NMR and cryo-electron microscopy is far from being negligible. Here, we summarize some highlights and illustrate the importance of specific technologies on the structural biology of protein-protein or protein/DNA transcription complexes: structure/ function analysis of components the eukaryotic basal and activated transcription machinery with focus on the TFIID and TFIIH multi-subunit complexes as well as transcription regulators such as members of the nuclear hormone receptor families. We also discuss molecular aspects of promoter recognition and epigenetic control of gene expression.

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Section: Challenges For Sample Preparation: New Methods For Protein Cmentioning

confidence: 99%

Structural insights into transcription complexes

Berger

Blanco

Boelens

et al. 2011

Journal of Structural Biology

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“…Recombinant production offers solutions to these impediments, and a wide range of expression systems are available to produce proteins recombinantly in prokaryotic and eukaryotic hosts [31,33,[34][35][36][37][38][39][40][41][42][43][44][45]. Recombinant expression systems share in common that one or several DNA segments encoding for proteins, protein domains or multicomponent protein complexes are typically combined with DNA elements including DNAs that control transcription (promoters, terminators, others) and translation (ribosome binding sites, ShineDalgarno sequences, Kozak consensus sequences, enhancers, others) and inserted into a functional DNA module (plasmid, cosmid, artificial chromosome, genome, others).…”

Section: / Acembl: Automated Unrestricted Dna Recombineering For Mulmentioning

confidence: 99%

ACEMBL Tool-Kits for High-Throughput Multigene Delivery and Expression in Prokaryotic and Eukaryotic Hosts

Nie

Chaillet

Becke

et al. 2016

Advances in Experimental Medicine and Biology

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“…Subunits of transient complexes may form crystals that exclude the other subunit, which is often difficult to detect. Recombinant production of the subunits of a protein complex in the same host cell by co-expression has been described with a large variety of systems (Busso et al 2011; Nie et al 2009; Vijayachandran et al 2011). Novel cloning strategies enable co-expression of many subunits in host cells including E. coli , baculovirus and mammalian cells (Berger et al 2004; Kriz et al 2010; Trowitzsch et al 2010), and have been adapted to automated cloning (Bieniossek et al 2009).…”

Section: Automation and Strategies For Protein Structure Analysismentioning

confidence: 99%

“…pQLink vectors have been widely used by different laboratories, mainly for eukaryotic vesicle tethering complexes (Kummel et al 2008; Lees et al 2010; Ren et al 2009). Studies comparing a large variety of expression systems have demonstrated that subtle changes in the expression strategy have a profound effect on the success of co-expression experiments, even if the main parameters, protein sequence and host cell are identical (Busso et al 2011). …”

Section: Automation and Strategies For Protein Structure Analysismentioning

confidence: 99%

Current methods in structural proteomics and its applications in biological sciences

et al. 2011

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A broad working definition of structural proteomics (SP) is that it is the process of the high-throughput characterization of the three-dimensional structures of biological macromolecules. Recently, the process for protein structure determination has become highly automated and SP platforms have been established around the globe, utilizing X-ray crystallography as a tool. Although protein structures often provide clues about the biological function of a target, once the three-dimensional structures have been determined, bioinformatics and proteomics-driven strategies can be employed to derive their biological activities and physiological roles. This article reviews the current status of SP methods for the structure determination pipeline, including target selection, isolation, expression, purification, crystallization, diffraction data collection, structure solution, refinement and functional annotation.

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Expression of protein complexes using multiple Escherichia coli protein co-expression systems: A benchmarking study

Cited by 38 publications

References 44 publications

Structural insights into transcription complexes

Structural insights into transcription complexes

ACEMBL Tool-Kits for High-Throughput Multigene Delivery and Expression in Prokaryotic and Eukaryotic Hosts

Current methods in structural proteomics and its applications in biological sciences

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