|
|
|
|
|
 |
Contents |
 |
|
|
|
|
|
|
|
|
|
|
|
|
* Residue conservation analysis
|
|
|
|
|
|
|
|
|
|
Enzyme class:
|
|
Chain T:
E.C.3.4.21.4
- trypsin.
|
|
|
|
|
|
|
Reaction:
|
|
Preferential cleavage: Arg-|-Xaa, Lys-|-Xaa.
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
 |
|
|
|
|
|
|
| |
|
DOI no:
|
J Biol Chem
272:19931-19937
(1997)
|
|
PubMed id:
|
|
|
|
|
|
| |
|
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.
|
|
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.
|
|
|
|
|
| |
ABSTRACT
|
|
|
|
| |
|
|
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.
|
|
|
|
|
|
| |
Selected figure(s)
|
|
|
|
| |
|
|
|
|
|
|
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).
|
|
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.
|
|
|
|
|
|
| |
The above figures are
reprinted
by permission from the ASBMB:
J Biol Chem
(1997,
272,
19931-19937)
copyright 1997.
|
|
| |
Figures were
selected
by an automated process.
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
 |
 |
 |
|
Literature references that cite this PDB file's key reference
|
|
|
| |
PubMed id
|
|
Reference
|
|
|
|
|
|
J.L.Arolas,
and
S.Ventura
(2011).
Protease inhibitors as models for the study of oxidative folding.
|
| |
Antioxid Redox Signal,
14,
97.
|
|
|
|
|
|
|
D.Pantoja-Uceda,
J.L.Arolas,
F.X.Aviles,
J.Santoro,
S.Ventura,
and
C.P.Sommerhoff
(2009).
Deciphering the structural basis that guides the oxidative folding of leech-derived tryptase inhibitor.
|
| |
J Biol Chem,
284,
35612-35620.
|
|
|
PDB codes:
|
|
|
|
|
|
|
|
M.Neira Oviedo,
J.M.Ribeiro,
A.Heyland,
L.VanEkeris,
T.Moroz,
and
P.J.Linser
(2009).
The salivary transcriptome of Anopheles gambiae (Diptera: Culicidae) larvae: A microarray-based analysis.
|
| |
Insect Biochem Mol Biol,
39,
382-394.
|
|
|
|
|
|
|
Y.Li,
Y.Q.Qian,
W.M.Ma,
and
W.J.Yang
(2009).
Inhibition mechanism and the effects of structure on activity of male reproduction-related peptidase inhibitor Kazal-type (MRPINK) of Macrobrachium rosenbergii.
|
| |
Mar Biotechnol (NY),
11,
252-259.
|
|
|
|
|
|
|
A.Iddamalgoda,
Q.T.Le,
K.Ito,
K.Tanaka,
H.Kojima,
and
H.Kido
(2008).
Mast cell tryptase and photoaging: possible involvement in the degradation of extra cellular matrix and basement membrane proteins.
|
| |
Arch Dermatol Res,
300,
S69-S76.
|
|
|
|
|
|
|
I.P.Baskova,
E.S.Kostrjukova,
M.A.Vlasova,
O.V.Kharitonova,
S.A.Levitskiy,
L.L.Zavalova,
S.A.Moshkovskii,
and
V.N.Lazarev
(2008).
Proteins and peptides of the salivary gland secretion of medicinal leeches Hirudo verbana, H. medicinalis, and H. orientalis.
|
| |
Biochemistry (Mosc),
73,
315-320.
|
|
|
|
|
|
|
J.L.Arolas,
S.Bronsoms,
F.X.Aviles,
S.Ventura,
and
C.P.Sommerhoff
(2008).
Oxidative folding of leech-derived tryptase inhibitor via native disulfide-bonded intermediates.
|
| |
Antioxid Redox Signal,
10,
77-86.
|
|
|
|
|
|
|
O.Avrutina,
H.U.Schmoldt,
D.Gabrijelcic-Geiger,
A.Wentzel,
H.Frauendorf,
C.P.Sommerhoff,
U.Diederichsen,
and
H.Kolmar
(2008).
Head-to-tail cyclized cystine-knot peptides by a combined recombinant and chemical route of synthesis.
|
| |
Chembiochem,
9,
33-37.
|
|
|
|
|
|
|
T.C.Assumpção,
I.M.Francischetti,
J.F.Andersen,
A.Schwarz,
J.M.Santana,
and
J.M.Ribeiro
(2008).
An insight into the sialome of the blood-sucking bug Triatoma infestans, a vector of Chagas' disease.
|
| |
Insect Biochem Mol Biol,
38,
213-232.
|
|
|
|
|
|
|
I.P.Baskova,
L.L.Zavalova,
E.S.Kostrjukova,
G.A.Titova,
V.N.Lazarev,
and
V.G.Zgoda
(2007).
Proteomic analysis methods for characterization of proteins from the salivary gland secretions of the medicinal leech during different seasons.
|
| |
Biochemistry (Mosc),
72,
219-225.
|
|
|
|
|
|
|
J.X.Cao,
J.Q.Dai,
Z.M.Dai,
G.L.Yin,
and
W.J.Yang
(2007).
A male reproduction-related Kazal-type peptidase inhibitor gene in the prawn, Macrobrachium rosenbergii: molecular characterization and expression patterns.
|
| |
Mar Biotechnol (NY),
9,
45-55.
|
|
|
|
|
|
|
Y.González,
T.Pons,
J.Gil,
V.Besada,
M.Alonso-del-Rivero,
A.S.Tanaka,
M.S.Araujo,
and
M.A.Chávez
(2007).
Characterization and comparative 3D modeling of CmPI-II, a novel 'non-classical' Kazal-type inhibitor from the marine snail Cenchritis muricatus (Mollusca).
|
| |
Biol Chem,
388,
1183-1194.
|
|
|
|
|
|
|
R.Krätzner,
J.E.Debreczeni,
T.Pape,
T.R.Schneider,
A.Wentzel,
H.Kolmar,
G.M.Sheldrick,
and
I.Uson
(2005).
Structure of Ecballium elaterium trypsin inhibitor II (EETI-II): a rigid molecular scaffold.
|
| |
Acta Crystallogr D Biol Crystallogr,
61,
1255-1262.
|
|
|
PDB codes:
|
|
|
|
|
|
|
|
A.Heifetz,
E.Katchalski-Katzir,
and
M.Eisenstein
(2002).
Electrostatics in protein-protein docking.
|
| |
Protein Sci,
11,
571-587.
|
|
|
|
|
|
|
D.Scarpi,
J.D.McBride,
and
R.J.Leatherbarrow
(2002).
Inhibition of human beta-tryptase by Bowman-Birk inhibitor derived peptides.
|
| |
J Pept Res,
59,
90-93.
|
|
|
|
|
|
|
H.Hemmi,
T.Yoshida,
T.Kumazaki,
N.Nemoto,
J.Hasegawa,
F.Nishioka,
Y.Kyogoku,
H.Yokosawa,
and
Y.Kobayashi
(2002).
Solution structure of ascidian trypsin inhibitor determined by nuclear magnetic resonance spectroscopy.
|
| |
Biochemistry,
41,
10657-10664.
|
|
|
PDB code:
|
|
|
|
|
|
|
|
F.Erba,
L.Fiorucci,
C.P.Sommerhoff,
M.Coletta,
and
F.Ascoli
(2000).
Kinetic and thermodynamic analysis of leech-derived tryptase inhibitor interaction with bovine tryptase and bovine trypsin.
|
| |
Biol Chem,
381,
1117-1122.
|
|
|
|
|
|
|
U.Rester,
M.Moser,
R.Huber,
and
W.Bode
(2000).
L-Isoaspartate 115 of porcine beta-trypsin promotes crystallization of its complex with bdellastasin.
|
| |
Acta Crystallogr D Biol Crystallogr,
56,
581-588.
|
|
|
PDB code:
|
|
|
|
|
|
|
|
C.P.Sommerhoff,
W.Bode,
P.J.Pereira,
M.T.Stubbs,
J.Stürzebecher,
G.P.Piechottka,
G.Matschiner,
and
A.Bergner
(1999).
The structure of the human betaII-tryptase tetramer: fo(u)r better or worse.
|
| |
Proc Natl Acad Sci U S A,
96,
10984-10991.
|
|
|
|
|
|
|
H.Czapinska,
and
J.Otlewski
(1999).
Structural and energetic determinants of the S1-site specificity in serine proteases.
|
| |
Eur J Biochem,
260,
571-595.
|
|
|
|
|
|
|
T.Niimi,
H.Yokoyama,
A.Goto,
K.Beck,
and
Y.Kitagawa
(1999).
A Drosophila gene encoding multiple splice variants of Kazal-type serine protease inhibitor-like proteins with potential destinations of mitochondria, cytosol and the secretory pathway.
|
| |
Eur J Biochem,
266,
282-292.
|
|
|
|
|
|
The most recent references are shown first.
Citation data come partly from CiteXplore and partly
from an automated harvesting procedure. Note that this is likely to be
only a partial list as not all journals are covered by
either method. However, we are continually building up the citation data
so more and more references will be included with time.
Where a reference describes a PDB structure, the PDB
codes are
shown on the right.
|
 |
Links
