1jcm
From Proteopedia
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| - | [[Image:1jcm.gif|left|200px]] | + | [[Image:1jcm.gif|left|200px]] |
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| - | '''TRPC STABILITY MUTANT CONTAINING AN ENGINEERED DISULPHIDE BRIDGE AND IN COMPLEX WITH A CDRP-RELATED SUBSTRATE''' | + | {{Structure |
| + | |PDB= 1jcm |SIZE=350|CAPTION= <scene name='initialview01'>1jcm</scene>, resolution 2.10Å | ||
| + | |SITE= | ||
| + | |LIGAND= <scene name='pdbligand=PO4:PHOSPHATE+ION'>PO4</scene> and <scene name='pdbligand=137:1-(O-CARBOXY-PHENYLAMINO)-1-DEOXY-D-RIBULOSE-5-PHOSPHATE'>137</scene> | ||
| + | |ACTIVITY= [http://en.wikipedia.org/wiki/Indole-3-glycerol-phosphate_synthase Indole-3-glycerol-phosphate synthase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=4.1.1.48 4.1.1.48] | ||
| + | |GENE= | ||
| + | }} | ||
| + | |||
| + | '''TRPC STABILITY MUTANT CONTAINING AN ENGINEERED DISULPHIDE BRIDGE AND IN COMPLEX WITH A CDRP-RELATED SUBSTRATE''' | ||
| + | |||
==Overview== | ==Overview== | ||
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==About this Structure== | ==About this Structure== | ||
| - | 1JCM is a [ | + | 1JCM is a [[Single protein]] structure of sequence from [http://en.wikipedia.org/wiki/Escherichia_coli Escherichia coli]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1JCM OCA]. |
==Reference== | ==Reference== | ||
| - | Stabilization of a (betaalpha)8-barrel protein by an engineered disulfide bridge., Ivens A, Mayans O, Szadkowski H, Jurgens C, Wilmanns M, Kirschner K, Eur J Biochem. 2002 Feb;269(4):1145-53. PMID:[http:// | + | Stabilization of a (betaalpha)8-barrel protein by an engineered disulfide bridge., Ivens A, Mayans O, Szadkowski H, Jurgens C, Wilmanns M, Kirschner K, Eur J Biochem. 2002 Feb;269(4):1145-53. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/11856350 11856350] |
[[Category: Escherichia coli]] | [[Category: Escherichia coli]] | ||
[[Category: Indole-3-glycerol-phosphate synthase]] | [[Category: Indole-3-glycerol-phosphate synthase]] | ||
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[[Category: stability mutant]] | [[Category: stability mutant]] | ||
| - | ''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 12:01:38 2008'' |
Revision as of 10:01, 20 March 2008
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| , resolution 2.10Å | |||||||
|---|---|---|---|---|---|---|---|
| Ligands: | and | ||||||
| Activity: | Indole-3-glycerol-phosphate synthase, with EC number 4.1.1.48 | ||||||
| Coordinates: | save as pdb, mmCIF, xml | ||||||
TRPC STABILITY MUTANT CONTAINING AN ENGINEERED DISULPHIDE BRIDGE AND IN COMPLEX WITH A CDRP-RELATED SUBSTRATE
Overview
The aim of this study was to increase the stability of the thermolabile (betaalpha)8-barrel enzyme indoleglycerol phosphate synthase from Escherichia coli by the introduction of disulfide bridges. For the design of such variants, we selected two out of 12 candidates, in which newly introduced cysteines potentially form optimal disulfide bonds. These variants avoid short-range connections, substitutions near catalytic residues, and crosslinks between the new and the three parental cysteines. The variant linking residues 3 and 189 fastens the N-terminus to the (betaalpha)8-barrel. The rate of thermal inactivation at 50 degrees C of this variant with a closed disulfide bridge is 65-fold slower than that of the reference dithiol form, but only 13-fold slower than that of the parental protein. The near-ultraviolet CD spectrum, the reactivity of parental buried cysteines with Ellman's reagent as well as the decreased turnover number indicate that the protein structure is rigidified. To confirm these data, we have solved the X-ray structure to 2.1-A resolution. The second variant was designed to crosslink the terminal modules betaalpha1 and betaalpha8. However, not even the dithiol form acquired the native fold, possibly because one of the targeted residues is solvent-inaccessible in the parental protein.
About this Structure
1JCM is a Single protein structure of sequence from Escherichia coli. Full crystallographic information is available from OCA.
Reference
Stabilization of a (betaalpha)8-barrel protein by an engineered disulfide bridge., Ivens A, Mayans O, Szadkowski H, Jurgens C, Wilmanns M, Kirschner K, Eur J Biochem. 2002 Feb;269(4):1145-53. PMID:11856350
Page seeded by OCA on Thu Mar 20 12:01:38 2008
