2cam
From Proteopedia
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| - | [[Image:2cam.gif|left|200px]] | + | [[Image:2cam.gif|left|200px]] |
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| - | '''AVIDIN MUTANT (K3E,K9E,R26D,R124L)''' | + | {{Structure |
| + | |PDB= 2cam |SIZE=350|CAPTION= <scene name='initialview01'>2cam</scene>, resolution 2.2Å | ||
| + | |SITE= <scene name='pdbsite=B1:Residues+Involved+In+Biotin+H+Bonding'>B1</scene> and <scene name='pdbsite=B2:Residues+Involved+In+Biotin+H+Bonding'>B2</scene> | ||
| + | |LIGAND= | ||
| + | |ACTIVITY= | ||
| + | |GENE= | ||
| + | }} | ||
| + | |||
| + | '''AVIDIN MUTANT (K3E,K9E,R26D,R124L)''' | ||
| + | |||
==Overview== | ==Overview== | ||
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==About this Structure== | ==About this Structure== | ||
| - | 2CAM is a [ | + | 2CAM 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=2CAM OCA]. |
==Reference== | ==Reference== | ||
| - | Biochemical characterization and crystal structure of a recombinant hen avidin and its acidic mutant expressed in Escherichia coli., Nardone E, Rosano C, Santambrogio P, Curnis F, Corti A, Magni F, Siccardi AG, Paganelli G, Losso R, Apreda B, Bolognesi M, Sidoli A, Arosio P, Eur J Biochem. 1998 Sep 1;256(2):453-60. PMID:[http:// | + | Biochemical characterization and crystal structure of a recombinant hen avidin and its acidic mutant expressed in Escherichia coli., Nardone E, Rosano C, Santambrogio P, Curnis F, Corti A, Magni F, Siccardi AG, Paganelli G, Losso R, Apreda B, Bolognesi M, Sidoli A, Arosio P, Eur J Biochem. 1998 Sep 1;256(2):453-60. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/9760187 9760187] |
[[Category: Gallus gallus]] | [[Category: Gallus gallus]] | ||
[[Category: Single protein]] | [[Category: Single protein]] | ||
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[[Category: avidin]] | [[Category: avidin]] | ||
[[Category: biotin binding protein]] | [[Category: biotin binding protein]] | ||
| - | [[Category: | + | [[Category: calycin]] |
[[Category: glycoprotein]] | [[Category: glycoprotein]] | ||
[[Category: up-and-down beta barrel]] | [[Category: up-and-down beta barrel]] | ||
| - | ''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 16:13:01 2008'' |
Revision as of 14:13, 20 March 2008
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| , resolution 2.2Å | |||||||
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| Sites: | and | ||||||
| Coordinates: | save as pdb, mmCIF, xml | ||||||
AVIDIN MUTANT (K3E,K9E,R26D,R124L)
Overview
The mature hen avidin encoded by a synthetic cDNA was expressed in Escherichia coli in an insoluble form. After resolubilization, renaturation and purification, a recovery of about 20 mg/l cell culture was obtained. ELISA assays indicated no apparent differences in biotin binding between the natural and recombinant avidins. In addition, an acidic avidin mutant, bearing the substitutions Lys3-->Glu, Lys9--> Glu, Arg26-->Asp and Arg124-->Leu of four exposed basic residues, was produced. The protein, expressed and renatured as wild-type avidin, showed unaltered biotin-binding activity. The acidic pI (approximately 5.5) and lack of aggregation of the mutant allowed easy electrophoretic analysis under non-denaturing conditions of the protein alone and of its complexes with biotin, biotinylated transferrin or peroxidase. Analysis of the sera from sensitized subjects revealed that the avidin mutant has altered antigenicity. Both recombinant avidins were crystallized and the three-dimensional structures solved by molecular replacement and refined to 0.22 nm resolution. The three-dimensional structures of the two recombinant molecules, in the absence of biotin and of glycosylation, are fully comparable with those of the natural hen avidin previously reported.
About this Structure
2CAM is a Single protein structure of sequence from Gallus gallus. Full crystallographic information is available from OCA.
Reference
Biochemical characterization and crystal structure of a recombinant hen avidin and its acidic mutant expressed in Escherichia coli., Nardone E, Rosano C, Santambrogio P, Curnis F, Corti A, Magni F, Siccardi AG, Paganelli G, Losso R, Apreda B, Bolognesi M, Sidoli A, Arosio P, Eur J Biochem. 1998 Sep 1;256(2):453-60. PMID:9760187
Page seeded by OCA on Thu Mar 20 16:13:01 2008
