1get: Difference between revisions
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'''ANATOMY OF AN ENGINEERED NAD-BINDING SITE''' | {{Structure | ||
|PDB= 1get |SIZE=350|CAPTION= <scene name='initialview01'>1get</scene>, resolution 2.00Å | |||
|SITE= | |||
|LIGAND= <scene name='pdbligand=FAD:FLAVIN-ADENINE+DINUCLEOTIDE'>FAD</scene> and <scene name='pdbligand=NAP:NADP NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE'>NAP</scene> | |||
|ACTIVITY= [http://en.wikipedia.org/wiki/Glutathione-disulfide_reductase Glutathione-disulfide reductase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.8.1.7 1.8.1.7] | |||
|GENE= | |||
}} | |||
'''ANATOMY OF AN ENGINEERED NAD-BINDING SITE''' | |||
==Overview== | ==Overview== | ||
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==About this Structure== | ==About this Structure== | ||
1GET is a [ | 1GET 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=1GET OCA]. | ||
==Reference== | ==Reference== | ||
Anatomy of an engineered NAD-binding site., Mittl PR, Berry A, Scrutton NS, Perham RN, Schulz GE, Protein Sci. 1994 Sep;3(9):1504-14. PMID:[http:// | Anatomy of an engineered NAD-binding site., Mittl PR, Berry A, Scrutton NS, Perham RN, Schulz GE, Protein Sci. 1994 Sep;3(9):1504-14. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/7833810 7833810] | ||
[[Category: Escherichia coli]] | [[Category: Escherichia coli]] | ||
[[Category: Glutathione-disulfide reductase]] | [[Category: Glutathione-disulfide reductase]] | ||
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[[Category: oxidoreductase(flavoenzyme)]] | [[Category: oxidoreductase(flavoenzyme)]] | ||
''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 11:22:04 2008'' | ||
Revision as of 12:22, 20 March 2008
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| , resolution 2.00Å | |||||||
|---|---|---|---|---|---|---|---|
| Ligands: | and | ||||||
| Activity: | Glutathione-disulfide reductase, with EC number 1.8.1.7 | ||||||
| Coordinates: | save as pdb, mmCIF, xml | ||||||
ANATOMY OF AN ENGINEERED NAD-BINDING SITE
OverviewOverview
The coenzyme specificity of Escherichia coli glutathione reductase was switched from NADP to NAD by modifying the environment of the 2'-phosphate binding site through a set of point mutations: A179G, A183G, V197E, R198M, K199F, H200D, and R204P (Scrutton NS, Berry A, Perham RN, 1990, Nature 343:38-43). In order to analyze the structural changes involved, we have determined 4 high-resolution crystal structures, i.e., the structures of the wild-type enzyme (1.86 A resolution, R-factor of 16.8%), of the wild-type enzyme ligated with NADP (2.0 A, 20.8%), of the NAD-dependent mutant (1.74 A, 16.8%), and of the NAD-dependent mutant ligated with NAD (2.2 A, 16.9%). A comparison of these structures reveals subtle differences that explain details of the specificity change. In particular, a peptide rotation occurs close to the adenosine ribose, with a concomitant change of the ribose pucker. The mutations cause a contraction of the local chain fold. Furthermore, the engineered NAD-binding site assumes a less rigid structure than the NADP site of the wild-type enzyme. A superposition of the ligated structures shows a displacement of NAD versus NADP such that the electron pathway from the nicotinamide ring to FAD is elongated, which may explain the lower catalytic efficiency of the mutant. Because the nicotinamide is as much as 15 A from the sites of the mutations, this observation reminds us that mutations may have important long-range consequences that are difficult to anticipate.
About this StructureAbout this Structure
1GET is a Single protein structure of sequence from Escherichia coli. Full crystallographic information is available from OCA.
ReferenceReference
Anatomy of an engineered NAD-binding site., Mittl PR, Berry A, Scrutton NS, Perham RN, Schulz GE, Protein Sci. 1994 Sep;3(9):1504-14. PMID:7833810
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