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PDBsum entry 1amu
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Peptide synthetase
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PDB id
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1amu
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Contents |
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* Residue conservation analysis
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References listed in PDB file
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Key reference
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Title
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Structural basis for the activation of phenylalanine in the non-Ribosomal biosynthesis of gramicidin s.
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Authors
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E.Conti,
T.Stachelhaus,
M.A.Marahiel,
P.Brick.
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Ref.
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Embo J, 1997,
16,
4174-4183.
[DOI no: ]
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PubMed id
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Abstract
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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.
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Figure 2.
Figure 2 Stereo diagram of the large N-terminal domain of PheA
showing the bound ligands coloured as in Figure 1. The  -sheet
A is on the left-hand side, -sheet
B is on the right-hand side, and the -barrel
is at the top of the figure. The N-terminus of the protein is at
the top of the figure. The disordered loop (residues 192 -196)
near the active site is coloured violet.
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Figure 6.
Figure 6 Schematic representation of the hydrogen bonding
between PheA and the phenylalanine and AMP ligands.
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The above figures are
reprinted
from an Open Access publication published by Macmillan Publishers Ltd:
Embo J
(1997,
16,
4174-4183)
copyright 1997.
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Secondary reference #1
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Title
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Modular structure of peptide synthetases revealed by dissection of the multifunctional enzyme grsa.
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Authors
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T.Stachelhaus,
M.A.Marahiel.
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Ref.
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J Biol Chem, 1995,
270,
6163-6169.
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PubMed id
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