High-throughput production of human proteins for crystallization: the SGC experience.
Producing purified human proteins with high yield and purity remains a considerable challenge. We describe the methods utilized in the Structural Genomics Consortium (SGC) in Oxford, resulting in successful purification of 48% of human proteins attempted; of those, the structures of approximately 40...
| Main Authors: | , , , , , , |
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| Format: | Journal article |
| Language: | English |
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2010
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| _version_ | 1797055089143185408 |
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| author | Savitsky, P Bray, J Cooper, C Marsden, B Mahajan, P Burgess-Brown, N Gileadi, O |
| author_facet | Savitsky, P Bray, J Cooper, C Marsden, B Mahajan, P Burgess-Brown, N Gileadi, O |
| author_sort | Savitsky, P |
| collection | OXFORD |
| description | Producing purified human proteins with high yield and purity remains a considerable challenge. We describe the methods utilized in the Structural Genomics Consortium (SGC) in Oxford, resulting in successful purification of 48% of human proteins attempted; of those, the structures of approximately 40% were solved by X-ray crystallography. The main driver has been the parallel processing of multiple (typically 9-20) truncated constructs of each target; modest diversity in vectors and host systems; and standardized purification procedures. We provide method details as well as data on the properties of the constructs leading to crystallized proteins and the impact of methodological variants. These can be used to formulate guidelines for initial approaches to expression of new eukaryotic proteins. |
| first_indexed | 2024-03-06T19:06:30Z |
| format | Journal article |
| id | oxford-uuid:1552adbd-3bae-402a-b875-b9437eb95ca7 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T19:06:30Z |
| publishDate | 2010 |
| record_format | dspace |
| spelling | oxford-uuid:1552adbd-3bae-402a-b875-b9437eb95ca72022-03-26T10:24:49ZHigh-throughput production of human proteins for crystallization: the SGC experience.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:1552adbd-3bae-402a-b875-b9437eb95ca7isolation and purificationchemistryCell LineProteomicsGenomicsHumansRecombinant ProteinsCrystallography, X-RayAmino Acid SequenceAnimalsmetabolismCloning, MoleculargeneticsmethodsGenetic VectorsMolecular Sequence DataProteinsEnglishStructural Genomics Consortium2010Savitsky, PBray, JCooper, CMarsden, BMahajan, PBurgess-Brown, NGileadi, OProducing purified human proteins with high yield and purity remains a considerable challenge. We describe the methods utilized in the Structural Genomics Consortium (SGC) in Oxford, resulting in successful purification of 48% of human proteins attempted; of those, the structures of approximately 40% were solved by X-ray crystallography. The main driver has been the parallel processing of multiple (typically 9-20) truncated constructs of each target; modest diversity in vectors and host systems; and standardized purification procedures. We provide method details as well as data on the properties of the constructs leading to crystallized proteins and the impact of methodological variants. These can be used to formulate guidelines for initial approaches to expression of new eukaryotic proteins. |
| spellingShingle | isolation and purification chemistry Cell Line Proteomics Genomics Humans Recombinant Proteins Crystallography, X-Ray Amino Acid Sequence Animals metabolism Cloning, Molecular genetics methods Genetic Vectors Molecular Sequence Data Proteins Savitsky, P Bray, J Cooper, C Marsden, B Mahajan, P Burgess-Brown, N Gileadi, O High-throughput production of human proteins for crystallization: the SGC experience. |
| title | High-throughput production of human proteins for crystallization: the SGC experience. |
| title_full | High-throughput production of human proteins for crystallization: the SGC experience. |
| title_fullStr | High-throughput production of human proteins for crystallization: the SGC experience. |
| title_full_unstemmed | High-throughput production of human proteins for crystallization: the SGC experience. |
| title_short | High-throughput production of human proteins for crystallization: the SGC experience. |
| title_sort | high throughput production of human proteins for crystallization the sgc experience |
| topic | isolation and purification chemistry Cell Line Proteomics Genomics Humans Recombinant Proteins Crystallography, X-Ray Amino Acid Sequence Animals metabolism Cloning, Molecular genetics methods Genetic Vectors Molecular Sequence Data Proteins |
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