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PDBsum entry 1h9l
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Hydrolase/hydrolase inhibitor
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PDB id
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1h9l
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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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Kinetic and crystallographic analysis of complexes formed between elastase and peptides from beta-Casein.
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Authors
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P.A.Wright,
R.C.Wilmouth,
I.J.Clifton,
C.J.Schofield.
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Ref.
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Eur J Biochem, 2001,
268,
2969-2974.
[DOI no: ]
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PubMed id
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Abstract
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Human beta-casomorphin-7 (NH2-Tyr-Pro-Phe-Val-Glu-Pro-Ile-CO2H) is a naturally
occurring peptide inhibitor of elastase that has been shown to form an
acyl-enzyme complex stable enough for X-ray crystallographic analysis at pH 5.
To investigate the importance of the N-terminal residues of the
beta-casomorphin-7 peptide for the inhibition of elastase, kinetic and
crystallographic analyses were undertaken to identify the minimum number of
residues required for effective formation of a stable complex between truncated
beta-casomorphin-7 peptides and porcine pancreatic elastase (PPE). The results
clearly demonstrate that significant inhibition of PPE can be effected by simple
tri-, tetra-and pentapeptides terminating in a carboxylic acid. These results
also suggest that in vivo regulation of protease activity could be mediated via
short peptides as well as by proteins. Crystallographic analysis of the complex
formed between N-acetyl-Val-Glu-Pro-Ile-CO2H and PPE at pH 5 (to 1.67 A
resolution) revealed an active site water molecule in an analogous position to
that observed in the PPE/beta-casomorphin-7 structure supportive of its
assignment as the 'hydrolytic water' in the deacylation step of serine protease
catalysis.
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Figure 1.
Fig. 1. Schematic diagram showing the antiparallel  sheet formed
between a productively bound peptide and the active site of PPE.
The nomenclature for the residues of the peptide (P[1]–P[4])
and their respective subsites in the active site (S[1]–S[4])
is also shown.
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Figure 2.
Fig. 2. Stereo-views of the acyl-enzyme complex formed
between N-acetyl-Val-Glu-Pro-Ile-CO[2]H (in gold) and PPE (in
green). The peptide is linked via an ester bond between its
C-terminal isoleucine and Ser195 of PPE. (A) Location of the
proposed hydrolytic water molecule (Wat532) hydrogen bonded to
N[  2] of His57
and poised for nucleophilic attack above the ester carbonyl. (B)
Antiparallel sheet formed
between N-acetyl-Val-Glu-Pro-Ile-CO[2]H and the Ser214–Val216
loop in the active site of PPE. The 2mF[o]DF[c] electron density
maps [26] were contoured at 1  . This figure
was produced using BOBSCRIPT [27], RASTER3D [28], IMAGEMAGICK
and PHOTOSHOP 5.0 (Adobe Systems Inc.).
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The above figures are
reprinted
by permission from the Federation of European Biochemical Societies:
Eur J Biochem
(2001,
268,
2969-2974)
copyright 2001.
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Secondary reference #1
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Title
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Structure of a specific acyl-Enzyme complex formed between beta-Casomorphin-7 and porcine pancreatic elastase.
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Authors
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R.C.Wilmouth,
I.J.Clifton,
C.V.Robinson,
P.L.Roach,
R.T.Aplin,
N.J.Westwood,
J.Hajdu,
C.J.Schofield.
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Ref.
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Nat Struct Biol, 1997,
4,
456-462.
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PubMed id
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Secondary reference #2
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Title
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Structure of native porcine pancreatic elastase at 1.65 a resolutions.
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Authors
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E.Meyer,
G.Cole,
R.Radhakrishnan,
O.Epp.
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Ref.
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Acta Crystallogr B, 1988,
44,
26-38.
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PubMed id
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Secondary reference #3
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Title
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'Ph-Jump' Crystallographic analyses of gamma-Lactam-Porcine pancreatic elastase complexes.
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Authors
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P.A.Wright,
R.C.Wilmouth,
I.J.Clifton,
C.J.Schofield.
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Ref.
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Biochem J, 2000,
351,
335-340.
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PubMed id
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