1jx9: Difference between revisions
New page: left|200px<br /><applet load="1jx9" size="450" color="white" frame="true" align="right" spinBox="true" caption="1jx9, resolution 2.28Å" /> '''Penicillin Acylase, ... |
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[[Image:1jx9.gif|left|200px]]<br /><applet load="1jx9" size=" | [[Image:1jx9.gif|left|200px]]<br /><applet load="1jx9" size="350" color="white" frame="true" align="right" spinBox="true" | ||
caption="1jx9, resolution 2.28Å" /> | caption="1jx9, resolution 2.28Å" /> | ||
'''Penicillin Acylase, mutant'''<br /> | '''Penicillin Acylase, mutant'''<br /> | ||
==Overview== | ==Overview== | ||
Penicillin acylase catalyses the condensation of Calpha-substituted | 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== | ==About this Structure== | ||
1JX9 is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Escherichia_coli Escherichia coli] with CA as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [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] Full crystallographic information is available from [http:// | 1JX9 is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Escherichia_coli Escherichia coli] with <scene name='pdbligand=CA:'>CA</scene> as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [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] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1JX9 OCA]. | ||
==Reference== | ==Reference== | ||
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[[Category: Penicillin amidase]] | [[Category: Penicillin amidase]] | ||
[[Category: Protein complex]] | [[Category: Protein complex]] | ||
[[Category: Dijkstra, B | [[Category: Dijkstra, B W.]] | ||
[[Category: Hensgens, C | [[Category: Hensgens, C M.H.]] | ||
[[Category: Keizer, E.]] | [[Category: Keizer, E.]] | ||
[[Category: Snijder, H | [[Category: Snijder, H J.]] | ||
[[Category: CA]] | [[Category: CA]] | ||
[[Category: beta-strands]] | [[Category: beta-strands]] | ||
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[[Category: ntn-hydrolase fold]] | [[Category: ntn-hydrolase fold]] | ||
''Page seeded by [http:// | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 13:27:43 2008'' | ||
Revision as of 14:27, 21 February 2008
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Penicillin Acylase, mutant
OverviewOverview
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 StructureAbout this Structure
1JX9 is a Protein complex structure of sequences from Escherichia coli with as ligand. Active as Penicillin amidase, with EC number 3.5.1.11 Full crystallographic information is available from OCA.
ReferenceReference
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
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