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PDBsum entry 1tcp
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Blood coagulation inhibitor
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PDB id
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1tcp
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DOI no:
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Protein Sci
4:178-186
(1995)
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PubMed id:
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NMR structure determination of tick anticoagulant peptide (TAP).
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M.S.Lim-Wilby,
K.Hallenga,
M.de Maeyer,
I.Lasters,
G.P.Vlasuk,
T.K.Brunck.
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ABSTRACT
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Tick anticoagulant peptide (TAP) is a potent and selective 60-amino acid
inhibitor of the serine protease Factor Xa (fXa), the penultimate enzyme in the
blood coagulation cascade. The structural features of TAP responsible for its
remarkable specificity for fXa are unknown, but the binding to its target
appears to be unique. The elucidation of the TAP structure may facilitate our
understanding of this new mode of serine protease inhibition and could provide a
basis for the design of novel fXa inhibitors. Analyses of homo- and
heteronuclear two-dimensional NMR spectra (total correlation spectroscopy,
nuclear Overhauser effect spectroscopy [NOESY], constant time heteronuclear
single quantum correlation spectroscopy [CT-HSQC], and HSQC-NOESY; 600 MHz; 1.5
mM TAP; pH 2.5) of unlabeled, 13C-labeled, and 15N-labeled TAP provided nearly
complete 1H sequence-specific resonance assignments. Secondary structural
elements were identified by characteristic NOE patterns and D2O amide
proton-exchange experiments. A three-dimensional structure of TAP was generated
from 412 NOESY-derived distance and 47 dihedral angle constraints. The
structural elements of TAP are similar in some respects to those of the Kunitz
serine protease inhibitor family, with which TAP shares weak sequence homology.
This structure, coupled with previous kinetic and biochemical information,
confirms previous suggestions that TAP has a unique mode of binding to fXa.
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Selected figure(s)
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Figure 1.
Fig. 1. Ca region f a'3C-constanttimeHSQCcorrelationexperi-
ment.Peaksare labeled by the single-letter codeandsequencenumber,
withthe Ca peaksdistinguishedfrom C@ andC6peaks by an enclos-
ing box.Theconstanttimeperiod was 17 ms,withatotal of 189 incre-
ments,eachasum of 64 transients.Processing was performedwith
TRIADsoftware(Tripos Associates). Theintensityofthe first point for
achID was halved, followed by apodization with asquared sine wave,
shifted by 80". Each FID was zero-filled to 2,048points,Fouriertrans-
formed,phased,andthen bascline correted with apolynomialfit.Pairs
ofinterferogramsfrom 512 incrementswereshuffled to separaereal
ndmaginaryparts.Thesecondtransform was perforedftersimi-
lar halvingofthefirstpointandapodization,followed by zerofilling
o 1,024 oints.Thespectrum was referenced to theTSP peak t 0 ppm
forbothcarbonandproton dimensions; two wl sweepwidths were added
to te ormer to obtainthewaleforthisregion.
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Figure 9.
Fig. 9. Superposition of thebakboneatoms
of arefinedTAPstructure(cyan)andBPTI
(yellow) (Marquartet al., 1983)alignedusing
backboneatoms (N, Ca, , 0) of residues22-
28,32-38, and51-60 of APwithbackbone
atoms of residues 18-24,29-35, and47-56 of
PTI,respectively.Selectedresiduenumbers
of TAP (1-4,30,42,60) andBPTI Ii 14,15,
8, 39, 58)are shown to provideanorienta-
tion. Cystinesidechainsaredepicted for each
olecule.Solvent-accessiblesurfaces(probe
adius 1.4 A) areshown as dotsurfacesforthe
riticalbindingresidues of TAP (1-4,42) and
BPTI (11, 13-19,34,36-39). Theorientation
isidentical to Figures6and 7.
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The above figures are
reprinted
from an Open Access publication published by the Protein Society:
Protein Sci
(1995,
4,
178-186)
copyright 1995.
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Figures were
selected
by an automated process.
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Literature references that cite this PDB file's key reference
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PubMed id
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Reference
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K.M.Bromfield,
N.S.Quinsey,
P.J.Duggan,
and
R.N.Pike
(2006).
Approaches to selective peptidic inhibitors of factor Xa.
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Chem Biol Drug Des,
68,
11-19.
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S.Salamanca,
and
J.Y.Chang
(2006).
Pathway of oxidative folding of a 3-disulfide alpha-lactalbumin may resemble either BPTI model or hirudin model.
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Protein J,
25,
275-287.
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R.St Charles,
K.Padmanabhan,
R.V.Arni,
K.P.Padmanabhan,
and
A.Tulinsky
(2000).
Structure of tick anticoagulant peptide at 1.6 A resolution complexed with bovine pancreatic trypsin inhibitor.
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Protein Sci,
9,
265-272.
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PDB code:
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A.Wei,
A.Smallwood,
R.S.Alexander,
J.Duke,
H.Ross,
S.A.Rosenfeld,
and
C.H.Chang
(1999).
Crystallization and preliminary X-ray diffraction data of the complex of recombinant tick anticoagulant peptide (rTAP) and bovine factor Xa.
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Acta Crystallogr D Biol Crystallogr,
55,
862-864.
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A.Sillen,
R.Vos,
and
Y.Engelborghs
(1996).
Fluorescence study of the conformational properties of recombinant tick anticoagulant peptide (Ornithodorus moubata) using multifrequency phase fluorometry.
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Photochem Photobiol,
64,
785-791.
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A.van de Locht,
M.T.Stubbs,
W.Bode,
T.Friedrich,
C.Bollschweiler,
W.Höffken,
and
R.Huber
(1996).
The ornithodorin-thrombin crystal structure, a key to the TAP enigma?
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EMBO J,
15,
6011-6017.
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PDB code:
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The most recent references are shown first.
Citation data come partly from CiteXplore and partly
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so more and more references will be included with time.
Where a reference describes a PDB structure, the PDB
code is
shown on the right.
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