1amu
From Proteopedia
(New page: 200px<br /><applet load="1amu" size="450" color="white" frame="true" align="right" spinBox="true" caption="1amu, resolution 1.9Å" /> '''PHENYLALANINE ACTIVAT...) |
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| - | [[Image:1amu.gif|left|200px]]<br /><applet load="1amu" size=" | + | [[Image:1amu.gif|left|200px]]<br /><applet load="1amu" size="350" color="white" frame="true" align="right" spinBox="true" |
caption="1amu, resolution 1.9Å" /> | caption="1amu, resolution 1.9Å" /> | ||
'''PHENYLALANINE ACTIVATING DOMAIN OF GRAMICIDIN SYNTHETASE 1 IN A COMPLEX WITH AMP AND PHENYLALANINE'''<br /> | '''PHENYLALANINE ACTIVATING DOMAIN OF GRAMICIDIN SYNTHETASE 1 IN A COMPLEX WITH AMP AND PHENYLALANINE'''<br /> | ||
==Overview== | ==Overview== | ||
| - | The non-ribosomal synthesis of the cyclic peptide antibiotic gramicidin S | + | The non-ribosomal synthesis of the cyclic peptide antibiotic gramicidin S is accomplished by two large multifunctional enzymes, the peptide synthetases 1 and 2. The enzyme complex contains five conserved subunits of approximately 60 kDa which carry out ATP-dependent activation of specific amino acids and share extensive regions of sequence similarity with adenylating enzymes such as firefly luciferases and acyl-CoA ligases. We have determined the crystal structure of the N-terminal adenylation subunit in a complex with AMP and L-phenylalanine to 1.9 A resolution. The 556 amino acid residue fragment is folded into two domains with the active site situated at their interface. Each domain of the enzyme has a similar topology to the corresponding domain of unliganded firefly luciferase, but a remarkable relative domain rotation of 94 degrees occurs. This conformation places the absolutely conserved Lys517 in a position to form electrostatic interactions with both ligands. The AMP is bound with the phosphate moiety interacting with Lys517 and the hydroxyl groups of the ribose forming hydrogen bonds with Asp413. The phenylalanine substrate binds in a hydrophobic pocket with the carboxylate group interacting with Lys517 and the alpha-amino group with Asp235. The structure reveals the role of the invariant residues within the superfamily of adenylate-forming enzymes and indicates a conserved mechanism of nucleotide binding and substrate activation. |
==About this Structure== | ==About this Structure== | ||
| - | 1AMU is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Brevibacillus_brevis Brevibacillus brevis] with MG, SO4, PHE and AMP as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Phenylalanine_racemase_(ATP-hydrolyzing) Phenylalanine racemase (ATP-hydrolyzing)], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=5.1.1.11 5.1.1.11] Full crystallographic information is available from [http:// | + | 1AMU is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Brevibacillus_brevis Brevibacillus brevis] with <scene name='pdbligand=MG:'>MG</scene>, <scene name='pdbligand=SO4:'>SO4</scene>, <scene name='pdbligand=PHE:'>PHE</scene> and <scene name='pdbligand=AMP:'>AMP</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Phenylalanine_racemase_(ATP-hydrolyzing) Phenylalanine racemase (ATP-hydrolyzing)], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=5.1.1.11 5.1.1.11] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1AMU OCA]. |
==Reference== | ==Reference== | ||
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[[Category: Brick, P.]] | [[Category: Brick, P.]] | ||
[[Category: Conti, E.]] | [[Category: Conti, E.]] | ||
| - | [[Category: Marahiel, M | + | [[Category: Marahiel, M A.]] |
[[Category: Stachelhaus, T.]] | [[Category: Stachelhaus, T.]] | ||
[[Category: AMP]] | [[Category: AMP]] | ||
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[[Category: peptide synthetase]] | [[Category: peptide synthetase]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 11:46:10 2008'' |
Revision as of 09:46, 21 February 2008
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PHENYLALANINE ACTIVATING DOMAIN OF GRAMICIDIN SYNTHETASE 1 IN A COMPLEX WITH AMP AND PHENYLALANINE
Overview
The non-ribosomal synthesis of the cyclic peptide antibiotic gramicidin S is accomplished by two large multifunctional enzymes, the peptide synthetases 1 and 2. The enzyme complex contains five conserved subunits of approximately 60 kDa which carry out ATP-dependent activation of specific amino acids and share extensive regions of sequence similarity with adenylating enzymes such as firefly luciferases and acyl-CoA ligases. We have determined the crystal structure of the N-terminal adenylation subunit in a complex with AMP and L-phenylalanine to 1.9 A resolution. The 556 amino acid residue fragment is folded into two domains with the active site situated at their interface. Each domain of the enzyme has a similar topology to the corresponding domain of unliganded firefly luciferase, but a remarkable relative domain rotation of 94 degrees occurs. This conformation places the absolutely conserved Lys517 in a position to form electrostatic interactions with both ligands. The AMP is bound with the phosphate moiety interacting with Lys517 and the hydroxyl groups of the ribose forming hydrogen bonds with Asp413. The phenylalanine substrate binds in a hydrophobic pocket with the carboxylate group interacting with Lys517 and the alpha-amino group with Asp235. The structure reveals the role of the invariant residues within the superfamily of adenylate-forming enzymes and indicates a conserved mechanism of nucleotide binding and substrate activation.
About this Structure
1AMU is a Single protein structure of sequence from Brevibacillus brevis with , , and as ligands. Active as Phenylalanine racemase (ATP-hydrolyzing), with EC number 5.1.1.11 Full crystallographic information is available from OCA.
Reference
Structural basis for the activation of phenylalanine in the non-ribosomal biosynthesis of gramicidin S., Conti E, Stachelhaus T, Marahiel MA, Brick P, EMBO J. 1997 Jul 16;16(14):4174-83. PMID:9250661
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