1kec
From Proteopedia
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| - | [[Image:1kec.jpg|left|200px]] | + | [[Image:1kec.jpg|left|200px]] |
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| - | '''PENICILLIN ACYLASE MUTANT WITH PHENYL PROPRIONIC ACID''' | + | {{Structure |
| + | |PDB= 1kec |SIZE=350|CAPTION= <scene name='initialview01'>1kec</scene>, resolution 2.30Å | ||
| + | |SITE= | ||
| + | |LIGAND= <scene name='pdbligand=CA:CALCIUM+ION'>CA</scene> and <scene name='pdbligand=GRO:R-2-PHENYL-PROPRIONIC ACID'>GRO</scene> | ||
| + | |ACTIVITY= [http://en.wikipedia.org/wiki/Penicillin_amidase Penicillin amidase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.5.1.11 3.5.1.11] | ||
| + | |GENE= PAC ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=562 Escherichia coli]) | ||
| + | }} | ||
| + | |||
| + | '''PENICILLIN ACYLASE MUTANT WITH PHENYL PROPRIONIC ACID''' | ||
| + | |||
==Overview== | ==Overview== | ||
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==About this Structure== | ==About this Structure== | ||
| - | 1KEC is a [ | + | 1KEC is a [[Protein complex]] structure of sequences 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=1KEC OCA]. |
==Reference== | ==Reference== | ||
| - | Structural and kinetic studies on ligand binding in wild-type and active-site mutants of penicillin acylase., Alkema WB, Hensgens CM, Snijder HJ, Keizer E, Dijkstra BW, Janssen DB, Protein Eng Des Sel. 2004 May;17(5):473-80. Epub 2004 Jul 14. PMID:[http:// | + | Structural and kinetic studies on ligand binding in wild-type and active-site mutants of penicillin acylase., Alkema WB, Hensgens CM, Snijder HJ, Keizer E, Dijkstra BW, Janssen DB, Protein Eng Des Sel. 2004 May;17(5):473-80. Epub 2004 Jul 14. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/15254299 15254299] |
[[Category: Escherichia coli]] | [[Category: Escherichia coli]] | ||
[[Category: Penicillin amidase]] | [[Category: Penicillin amidase]] | ||
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[[Category: CA]] | [[Category: CA]] | ||
[[Category: GRO]] | [[Category: GRO]] | ||
| - | [[Category: beta- | + | [[Category: beta-strand]] |
| - | [[Category: | + | [[Category: helice]] |
[[Category: ntn-hydrolase fold]] | [[Category: ntn-hydrolase fold]] | ||
[[Category: phenyl proprionic acid]] | [[Category: phenyl proprionic acid]] | ||
| - | ''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 12:16:16 2008'' |
Revision as of 10:16, 20 March 2008
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| , resolution 2.30Å | |||||||
|---|---|---|---|---|---|---|---|
| Ligands: | and | ||||||
| Gene: | PAC (Escherichia coli) | ||||||
| Activity: | Penicillin amidase, with EC number 3.5.1.11 | ||||||
| Coordinates: | save as pdb, mmCIF, xml | ||||||
PENICILLIN ACYLASE MUTANT WITH PHENYL PROPRIONIC ACID
Overview
Penicillin acylase catalyses the condensation of Calpha-substituted phenylacetic acids with beta-lactam nucleophiles, producing semi-synthetic beta-lactam antibiotics. For efficient synthesis a low affinity for phenylacetic acid and a high affinity for Calpha-substituted phenylacetic acid derivatives is desirable. We made three active site mutants, alphaF146Y, betaF24A and alphaF146Y/betaF24A, which all had a 2- to 10-fold higher affinity for Calpha-substituted compounds than wild-type enzyme. In addition, betaF24A had a 20-fold reduced affinity for phenylacetic acid. The molecular basis of the improved properties was investigated by X-ray crystallography. These studies showed that the higher affinity of alphaF146Y for (R)-alpha-methylphenylacetic acid can be explained by van der Waals interactions between alphaY146:OH and the Calpha-substituent. The betaF24A mutation causes an opening of the phenylacetic acid binding site. Only (R)-alpha-methylphenylacetic acid, but not phenylacetic acid, induces a conformation with the ligand tightly bound, explaining the weak binding of phenylacetic acid. A comparison of the betaF24A structure with other open conformations of penicillin acylase showed that betaF24 has a fixed position, whereas alphaF146 acts as a flexible lid on the binding site and reorients its position to achieve optimal substrate binding.
About this Structure
1KEC is a Protein complex structure of sequences from Escherichia coli. Full crystallographic information is available from OCA.
Reference
Structural and kinetic studies on ligand binding in wild-type and active-site mutants of penicillin acylase., Alkema WB, Hensgens CM, Snijder HJ, Keizer E, Dijkstra BW, Janssen DB, Protein Eng Des Sel. 2004 May;17(5):473-80. Epub 2004 Jul 14. PMID:15254299
Page seeded by OCA on Thu Mar 20 12:16:16 2008
