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| - | [[Image:2bsw.gif|left|200px]] | + | {{Seed}} |
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| | {{STRUCTURE_2bsw| PDB=2bsw | SCENE= }} | | {{STRUCTURE_2bsw| PDB=2bsw | SCENE= }} |
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| - | '''CRYSTAL STRUCTURE OF A GLYPHOSATE-N-ACETYLTRANSFERASE OBTAINED BY DNA SHUFFLING.'''
| + | ===CRYSTAL STRUCTURE OF A GLYPHOSATE-N-ACETYLTRANSFERASE OBTAINED BY DNA SHUFFLING.=== |
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| - | ==Overview==
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| - | The success of structural studies performed on an individual target in small scale or on many targets in the system-wide scale of structural genomics depends critically on three parameters: (i) obtaining an expression system capable of producing large quantities of the macromolecule(s) of interest, (ii) purifying this material in soluble form, and (iii) obtaining diffraction-quality crystals suitable for x-ray analysis. The attrition rate caused by these constraints is often quite high. Here, we present a strategy that addresses each of these three parameters simultaneously. Using DNA shuffling to introduce functional sequence variability into a protein of interest, we screened crude lysate supernatants for soluble variants that retain enzymatic activity. Crystallization trials performed on three WT and eight shuffled enzymes revealed two variants that crystallized readily. One of these was used to determine the high-resolution structure of the enzyme by x-ray analysis. The sequence diversity introduced through shuffling efficiently samples crystal packing space by modifying the surface properties of the enzyme. The approach demonstrated here does not require guidance as to the type of mutation necessary for improvements in expression, solubility, or crystallization. The method is scaleable and can be applied in situations where a single protein is being studied or in high-throughput structural genomics programs. Furthermore, it should be readily applied to structural studies of soluble proteins, membrane proteins, and macromolecular complexes. | + | The line below this paragraph, {{ABSTRACT_PUBMED_15951425}}, adds the Publication Abstract to the page |
| | + | (as it appears on PubMed at http://www.pubmed.gov), where 15951425 is the PubMed ID number. |
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| | + | {{ABSTRACT_PUBMED_15951425}} |
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| | ==About this Structure== | | ==About this Structure== |
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| | [[Category: Gnat superfamily]] | | [[Category: Gnat superfamily]] |
| | [[Category: Transferase]] | | [[Category: Transferase]] |
| - | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sat May 3 20:45:11 2008'' | + | |
| | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Tue Jul 29 14:41:12 2008'' |
Revision as of 11:41, 29 July 2008
Template:STRUCTURE 2bsw
CRYSTAL STRUCTURE OF A GLYPHOSATE-N-ACETYLTRANSFERASE OBTAINED BY DNA SHUFFLING.
Template:ABSTRACT PUBMED 15951425
About this Structure
2BSW is a Single protein structure. Full crystallographic information is available from OCA.
Reference
DNA shuffling as a tool for protein crystallization., Keenan RJ, Siehl DL, Gorton R, Castle LA, Proc Natl Acad Sci U S A. 2005 Jun 21;102(25):8887-92. Epub 2005 Jun 10. PMID:15951425
Page seeded by OCA on Tue Jul 29 14:41:12 2008
