 |
PDBsum entry 3ghn
|
|
|
|
References listed in PDB file
|
 |
|
Key reference
|
|
|
Title
|
|
Crystal structures of the noncatalytic domains of adamts13 reveal multiple discontinuous exosites for von willebrand factor.
|
|
|
Authors
|
|
M.Akiyama,
S.Takeda,
K.Kokame,
J.Takagi,
T.Miyata.
|
|
|
Ref.
|
|
Proc Natl Acad Sci U S A, 2009,
106,
19274-19279.
[DOI no: ]
|
|
|
PubMed id
|
|
|
|
|
Note In the PDB file this reference is
annotated as "TO BE PUBLISHED".
The citation details given above were identified by an automated
search of PubMed on title and author
names, giving a
perfect match.
|
|
|
|
Abstract
|
|
|
ADAMTS13 specifically cleaves plasma von Willebrand factor (VWF) and thereby
controls VWF-mediated platelet thrombus formation. Severe deficiencies in
ADAMTS13 can cause life-threatening thrombotic thrombocytopenic purpura. Here,
we determined 2 crystal structures of ADAMTS13-DTCS (residues 287-685), an
exosite-containing human ADAMTS13 fragment, at 2.6-A and 2.8-A resolution. The
structures revealed folding similarities between the disintegrin-like (D) domain
and the N-terminal portion of the cysteine-rich domain (designated the C(A)
domain). The spacer (S) domain forms a globular functional unit with a
10-stranded beta-sandwich fold that has multiple interaction sites with the C(A)
domain. We expressed 25 structure-based mutants of ADAMTS13-MDTCS (residues
75-685) and measured their enzymatic activity. We identified 3 VWF-binding
exosites on the linearly aligned discontinuous surfaces of the D, C(A), and S
domains traversing the W-shaped molecule. Since the MDTCS domains are conserved
among ADAMTS family proteins, the structural framework of the multiple
enzyme-substrate interactions identified in the ADAMTS13-VWF system provides the
basis for a common substrate recognition mode in this class of proteinases.
|
|
|
|
|
|
|
|
Figure 1.
Structure of ADAMTS13-DTCS. (A) Schematic representation of
the domain structures of full-length ADAMTS13 and ADAMTS13-DTCS.
(B) Ribbon structure of ADAMTS13-DTCS (form-1) in stereo.
Domains are colored as in A. Strands in the S domain are
numbered.
|
|
Figure 4.
ADAMTS13-VWF interactions. (A) Folded and unfolded structures
of the VWF A2 domain. The VWF A2 domain adopts a Rossman fold
with a central 6-stranded β-sheet surrounded by 5 α-helices
(shown as “A2 folded”) (28). The scissile peptide bond
(Tyr-1605-Met-1606) is buried within the protein core under
static conditions. The C-terminal region (residues
1,596–1,668, corresponding to VWF73) (31) of the A2 domain
must be unfolded to expose the scissile bond and the
exosite-binding regions under shear-stress conditions (shown as
A2 unfolded). (B) ADAMTS13-MDTCS-VWF binding model. The
molecular surface of the ADAMTS13-MDTCS model is shown in gray
and the bound zinc ion is shown in yellow. Residues that mediate
VWF binding are depicted as in Fig. 3C, and the exosites and the
catalytic cleft are indicated by red and yellow dotted
ellipsoids, respectively. The dotted green line represents a VWF
molecule (residues 1,596–1,668) bound to ADAMTS-MDTCS. (C)
Close-up view of the α6 helix and surrounding residues in the
VWF A2 domain. Hydrophobic residues are indicated with red
letters. Systematic charge-to-alanine substitutions revealed
that the D1653A and D1663A mutations (cyan) reduced the
substrate cleavage, the E1655A mutation (orange) slightly
increased cleavage, and the R1659A, E1660A, and R1668A mutations
(gray) had no significant effect (34).
|
|
|
|
|
|
Secondary reference #1
|
|
|
Title
|
|
Production, Crystallization and preliminary crystallographic analysis of an exosite-Containing fragment of human von willebrand factor-Cleaving proteinase adamts13.
|
|
|
Authors
|
|
M.Akiyama,
S.Takeda,
K.Kokame,
J.Takagi,
T.Miyata.
|
|
|
Ref.
|
|
Acta Crystallogr Sect F Struct Biol Cryst Commun, 2009,
65,
739-742.
|
|
|
PubMed id
|
|
|
|
|
|
|
|
|
|
|
|
|
