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PDBsum entry 1ldt
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Complex (hydrolase/inhibitor)
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PDB id
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1ldt
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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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The three-Dimensional structure of recombinant leech-Derived tryptase inhibitor in complex with trypsin. Implications for the structure of human mast cell tryptase and its inhibition.
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Authors
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M.T.Stubbs,
R.Morenweiser,
J.Stürzebecher,
M.Bauer,
W.Bode,
R.Huber,
G.P.Piechottka,
G.Matschiner,
C.P.Sommerhoff,
H.Fritz,
E.A.Auerswald.
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Ref.
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J Biol Chem, 1997,
272,
19931-19937.
[DOI no: ]
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PubMed id
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Abstract
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The x-ray crystal structure of recombinant leech-derived tryptase inhibitor
(rLDTI) has been solved to a resolution of 1.9 A in complex with porcine
trypsin. The nonclassical Kazal-type inhibitor exhibits the same overall
architecture as that observed in solution and in rhodniin. The complex reveals
structural aspects of the mast cell proteinase tryptase. The conformation of the
binding region of rLDTI suggests that tryptase has a restricted active site
cleft. The basic amino terminus of rLDTI, apparently flexible from previous NMR
measurements, approaches the 148-loop of trypsin. This loop has an acidic
equivalent in tryptase, suggesting that the basic amino terminus could make
favorable electrostatic interactions with the tryptase molecule. A series of
rLDTI variants constructed to probe this hypothesis confirmed that the
amino-terminal Lys-Lys sequence plays a role in inhibition of human lung
tryptase but not of trypsin or chymotrypsin. The location of such an acidic
surface patch is in accordance with the known low molecular weight inhibitors of
tryptase.
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Figure 1.
Fig. 1. Experimental electron density in the vicinity of the
active site in standard orientation (see Fig. 3). The binding of
LDTI (green) to trypsin (orange) causes the side chain of Tyr217
to flip outwards. This is a result of the conformation of the^
disulfide Cys4I-Cys29I ("I" suffix used to distinguish inhibitor
residue numbers from those of trypsin). This figure was prepared
using the program O (30).
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Figure 3.
Fig. 3. Schematic road map of human tryptase. A, view of
LDTI's binding loop (green sticks) in the active site cleft of
trypsin (orange sticks), displayed together with trypsin's
Connolly dot surface (figure prepared using MAIN (58)). Loops
contributing to the border of the active site are labeled. B,
sequence of corresponding loops of human tryptase in the
vicinity of the active site (white^ triangle). Green circles,
hydrophobic residues; red, acidic residues; blue, basic
residues; light blue, other polar residues. The positions of two
large insertions with respect to trypsin are shaded. The^
9-residue insertion between trypsin residues 175 and 176 would^
occlude the active site of tryptase to the west. The acidic
148-loop could receive the basic amino terminus of LDTI and
basic groups of low molecular weight inhibitors. See
"Discussion" for details.
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The above figures are
reprinted
by permission from the ASBMB:
J Biol Chem
(1997,
272,
19931-19937)
copyright 1997.
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Secondary reference #1
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Title
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Structure of leech derived tryptase inhibitor (ldti-C) in solution.
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Authors
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P.Mühlhahn,
M.Czisch,
R.Morenweiser,
B.Habermann,
R.A.Engh,
C.P.Sommerhoff,
E.A.Auerswald,
T.A.Holak.
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Ref.
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FEBS Lett, 1994,
355,
290-296.
[DOI no: ]
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PubMed id
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Secondary reference #2
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Title
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Recombinant leech-Derived tryptase inhibitor: construction, Production, Protein chemical characterization and inhibition of HIV-1 replication.
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Authors
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E.A.Auerswald,
R.Morenweiser,
C.P.Sommerhoff,
G.P.Piechottka,
C.Eckerskorn,
L.G.Gürtler,
H.Fritz.
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Ref.
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Biol Chem Hoppe Seyler, 1994,
375,
695-703.
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PubMed id
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Secondary reference #3
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Title
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A kazal-Type inhibitor of human mast cell tryptase: isolation from the medical leech hirudo medicinalis, Characterization, And sequence analysis.
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Authors
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C.P.Sommerhoff,
C.Söllner,
R.Mentele,
G.P.Piechottka,
E.A.Auerswald,
H.Fritz.
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Ref.
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Biol Chem Hoppe Seyler, 1994,
375,
685-694.
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PubMed id
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