Expressing the human proteome for affinity proteomics

Optimising expression of soluble protein domains and in vivo biotinylation

Tracy Keates, Christopher D O Cooper, Pavel Savitsky, Charles K. Allerston, Claire Phillips, Martin Hammarström, Neha Daga, Georgina Berridge, Pravin Mahajan, Nicola A. Burgess-Brown, Susanne Müller, Susanne Gräslund, Opher Gileadi

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

The generation of affinity reagents to large numbers of human proteins depends on the ability to express the target proteins as high-quality antigens. The Structural Genomics Consortium (SGC) focuses on the production and structure determination of human proteins. In a 7-year period, the SGC has deposited crystal structures of >800 human protein domains, and has additionally expressed and purified a similar number of protein domains that have not yet been crystallised. The targets include a diversity of protein domains, with an attempt to provide high coverage of protein families. The family approach provides an excellent basis for characterising the selectivity of affinity reagents. We present a summary of the approaches used to generate purified human proteins or protein domains, a test case demonstrating the ability to rapidly generate new proteins, and an optimisation study on the modification of >70 proteins by biotinylation in vivo. These results provide a unique synergy between large-scale structural projects and the recent efforts to produce a wide coverage of affinity reagents to the human proteome.

Original languageEnglish
Pages (from-to)515-525
Number of pages11
JournalNew Biotechnology
Volume29
Issue number5
DOIs
Publication statusPublished - 15 Jun 2012
Externally publishedYes

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Biotinylation
Proteome
Proteomics
Proteins
Genomics
Protein Domains
Antigens
Crystal structure

Cite this

Keates, Tracy ; Cooper, Christopher D O ; Savitsky, Pavel ; Allerston, Charles K. ; Phillips, Claire ; Hammarström, Martin ; Daga, Neha ; Berridge, Georgina ; Mahajan, Pravin ; Burgess-Brown, Nicola A. ; Müller, Susanne ; Gräslund, Susanne ; Gileadi, Opher. / Expressing the human proteome for affinity proteomics : Optimising expression of soluble protein domains and in vivo biotinylation. In: New Biotechnology. 2012 ; Vol. 29, No. 5. pp. 515-525.
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Keates, T, Cooper, CDO, Savitsky, P, Allerston, CK, Phillips, C, Hammarström, M, Daga, N, Berridge, G, Mahajan, P, Burgess-Brown, NA, Müller, S, Gräslund, S & Gileadi, O 2012, 'Expressing the human proteome for affinity proteomics: Optimising expression of soluble protein domains and in vivo biotinylation', New Biotechnology, vol. 29, no. 5, pp. 515-525. https://doi.org/10.1016/j.nbt.2011.10.007

Expressing the human proteome for affinity proteomics : Optimising expression of soluble protein domains and in vivo biotinylation. / Keates, Tracy; Cooper, Christopher D O; Savitsky, Pavel; Allerston, Charles K.; Phillips, Claire; Hammarström, Martin; Daga, Neha; Berridge, Georgina; Mahajan, Pravin; Burgess-Brown, Nicola A.; Müller, Susanne; Gräslund, Susanne; Gileadi, Opher.

In: New Biotechnology, Vol. 29, No. 5, 15.06.2012, p. 515-525.

Research output: Contribution to journalArticle

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AU - Gileadi, Opher

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