Expressing the human proteome for affinity proteomics: optimising expression of soluble protein domains and in vivo biotinylation.
N Biotechnol
; 29(5): 515-25, 2012 Jun 15.
Article
in En
| MEDLINE
| ID: mdl-22027370
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.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Chromatography, Affinity
/
Proteome
/
Proteomics
Limits:
Animals
/
Humans
Language:
En
Journal:
N Biotechnol
Journal subject:
BIOLOGIA MOLECULAR
/
ENGENHARIA BIOMEDICA
Year:
2012
Document type:
Article
Affiliation country: