1neg
From Proteopedia
| Line 1: | Line 1: | ||
| - | [[Image:1neg.gif|left|200px]] | + | [[Image:1neg.gif|left|200px]] |
| - | + | ||
| - | '''Crystal Structure Analysis of N-and C-terminal labeled SH3-domain of alpha-Chicken Spectrin''' | + | {{Structure |
| + | |PDB= 1neg |SIZE=350|CAPTION= <scene name='initialview01'>1neg</scene>, resolution 2.30Å | ||
| + | |SITE= | ||
| + | |LIGAND= <scene name='pdbligand=AZI:AZIDE ION'>AZI</scene> | ||
| + | |ACTIVITY= | ||
| + | |GENE= | ||
| + | }} | ||
| + | |||
| + | '''Crystal Structure Analysis of N-and C-terminal labeled SH3-domain of alpha-Chicken Spectrin''' | ||
| + | |||
==Overview== | ==Overview== | ||
| Line 7: | Line 16: | ||
==About this Structure== | ==About this Structure== | ||
| - | 1NEG is a [ | + | 1NEG is a [[Single protein]] structure of sequence from [http://en.wikipedia.org/wiki/Gallus_gallus Gallus gallus]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1NEG OCA]. |
==Reference== | ==Reference== | ||
| - | Rapid purification and crystal structure analysis of a small protein carrying two terminal affinity tags., Mueller U, Bussow K, Diehl A, Bartl FJ, Niesen FH, Nyarsik L, Heinemann U, J Struct Funct Genomics. 2003;4(4):217-25. PMID:[http:// | + | Rapid purification and crystal structure analysis of a small protein carrying two terminal affinity tags., Mueller U, Bussow K, Diehl A, Bartl FJ, Niesen FH, Nyarsik L, Heinemann U, J Struct Funct Genomics. 2003;4(4):217-25. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/15185962 15185962] |
[[Category: Gallus gallus]] | [[Category: Gallus gallus]] | ||
[[Category: Single protein]] | [[Category: Single protein]] | ||
| Line 20: | Line 29: | ||
[[Category: Nyarsik, L.]] | [[Category: Nyarsik, L.]] | ||
[[Category: AZI]] | [[Category: AZI]] | ||
| - | [[Category: five antiparallel beta | + | [[Category: five antiparallel beta sheet]] |
[[Category: sh3-domain fold]] | [[Category: sh3-domain fold]] | ||
| - | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Mar 20 12:55:47 2008'' |
Revision as of 10:55, 20 March 2008
| |||||||
| , resolution 2.30Å | |||||||
|---|---|---|---|---|---|---|---|
| Ligands: | |||||||
| Coordinates: | save as pdb, mmCIF, xml | ||||||
Crystal Structure Analysis of N-and C-terminal labeled SH3-domain of alpha-Chicken Spectrin
Overview
Small peptide tags are often fused to proteins to allow their affinity purification in high-throughput structure analysis schemes. To assess the compatibility of small peptide tags with protein crystallization and to examine if the tags alter the three-dimensional structure, the N-terminus of the chicken alpha-spectrin SH3 domain was labeled with a His6 tag and the C-terminus with a StrepII tag. The resulting protein, His6-SH3-StrepII, consists of 83 amino-acid residues, 23 of which originate from the tags. His6-SH3-StrepII is readily purified by dual affinity chromatography, has very similar biophysical characteristics as the untagged protein domain and crystallizes readily from a number of sparse-matrix screen conditions. The crystal structure analysis at 2.3 A resolution proves native-like structure of His6-SH3-StrepII and shows the entire His6 tag and part of the StrepII tag to be disordered in the crystal. Obviously, the fused affinity tags did not interfere with crystallization and structure analysis and did not change the protein structure. From the extreme case of His6-SH3-StrepII, where affinity tags represent 27% of the total fusion protein mass, we extrapolate that protein constructs with N- and C-terminal peptide tags may lend themselves to biophysical and structural investigations in high-throughput regimes.
About this Structure
1NEG is a Single protein structure of sequence from Gallus gallus. Full crystallographic information is available from OCA.
Reference
Rapid purification and crystal structure analysis of a small protein carrying two terminal affinity tags., Mueller U, Bussow K, Diehl A, Bartl FJ, Niesen FH, Nyarsik L, Heinemann U, J Struct Funct Genomics. 2003;4(4):217-25. PMID:15185962
Page seeded by OCA on Thu Mar 20 12:55:47 2008
