TY - JOUR
T1 - Expressing the human proteome for affinity proteomics
T2 - Optimising expression of soluble protein domains and in vivo biotinylation
AU - Keates, Tracy
AU - Cooper, Christopher D O
AU - Savitsky, Pavel
AU - Allerston, Charles K.
AU - Phillips, Claire
AU - Hammarström, Martin
AU - Daga, Neha
AU - Berridge, Georgina
AU - Mahajan, Pravin
AU - Burgess-Brown, Nicola A.
AU - Müller, Susanne
AU - Gräslund, Susanne
AU - Gileadi, Opher
PY - 2012/6/15
Y1 - 2012/6/15
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=84862024555&partnerID=8YFLogxK
UR - https://www.journals.elsevier.com/new-biotechnology/
U2 - 10.1016/j.nbt.2011.10.007
DO - 10.1016/j.nbt.2011.10.007
M3 - Article
C2 - 22027370
AN - SCOPUS:84862024555
VL - 29
SP - 515
EP - 525
JO - Biomolecular Engineering
JF - Biomolecular Engineering
SN - 1871-6784
IS - 5
ER -