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PMID: 19620974 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S.

Robust, high-throughput solution structural analyses by small angle X-ray scattering (SAXS).

Nature methods ·Vol. 6 ·No. 8 ·2009-08-00 ·Pages 606-12

Hura GL, Menon AL, Hammel M, Rambo RP, Poole FL, Tsutakawa SE, Jenney FE, Classen S, Frankel KA, Hopkins RC, Yang SJ, Scott JW, Dillard BD, Adams MW, Tainer JA

Abstract

We present an efficient pipeline enabling high-throughput analysis of protein structure in solution with small angle X-ray scattering (SAXS). Our SAXS pipeline combines automated sample handling of microliter volumes, temperature and anaerobic control, rapid data collection and data analysis, and couples structural analysis with automated archiving. We subjected 50 representative proteins, mostly from Pyrococcus furiosus, to this pipeline and found that 30 were multimeric structures in solution. SAXS analysis allowed us to distinguish aggregated and unfolded proteins, define global structural parameters and oligomeric states for most samples, identify shapes and similar structures for 25 unknown structures, and determine envelopes for 41 proteins. We believe that high-throughput SAXS is an enabling technology that may change the way that structural genomics research is done.

MeSH Terms
Bacterial Proteins/chemistry Equipment Design Models, Molecular Protein Conformation Proteins/chemistry Pyrococcus furiosus/metabolism Scattering, Small Angle X-Ray Diffraction/instrumentation,methods
Chemicals
Bacterial Proteins Proteins
Authors & Affiliations
15 authors, click to expand affiliations / ORCID
Hura Greg L
Physical Bioscience Division, Lawrence Berkeley National Laboratory, Berkeley, California, USA.
Menon Angeli L
Hammel Michal
Rambo Robert P
Poole Farris L
Tsutakawa Susan E
Jenney Francis E
Classen Scott
Frankel Kenneth A
Hopkins Robert C
Yang Sung-Jae
Scott Joseph W
Dillard Bret D
Adams Michael W W
Tainer John A
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Article Info
Journal
Nature methods
Abbr.
Nat Methods
ISSN
1548-7105
DOI
10.1038/nmeth.1353
Published
2009-08-00
Epub
2009-00-20
Pages
606-12
Language
English
Region
United States
NLM ID
101215604
PMCID
PMC3094553
Subset
IM
Grants
NCI NIH HHS · R01 CA117638 · United States
NCI NIH HHS · R01 CA117638-06 · United States
NCI NIH HHS · R01 CA117638-07 · United States
NCI NIH HHS · R01 CA117638-08 · United States
External Links
PubMed source DOI: 10.1038/nmeth.1353 PMC Full Text View journal details
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