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PDBsum entry 2v9t
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Structural protein/receptor
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PDB id
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2v9t
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References listed in PDB file
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Key reference
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Title
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Structural insights into the slit-Robo complex.
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Authors
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C.Morlot,
N.M.Thielens,
R.B.Ravelli,
W.Hemrika,
R.A.Romijn,
P.Gros,
S.Cusack,
A.A.Mccarthy.
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Ref.
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Proc Natl Acad Sci U S A, 2007,
104,
14923-14928.
[DOI no: ]
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PubMed id
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Abstract
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Slits are large multidomain leucine-rich repeat (LRR)-containing proteins that
provide crucial guidance cues in neuronal and vascular development. More
recently, Slits have been implicated in heart morphogenesis, angiogenesis, and
tumor metastasis. Slits are ligands for the Robo (Roundabout) receptors, which
belong to the Ig superfamily of transmembrane signaling molecules. The Slit-Robo
interaction is mediated by the second LRR domain of Slit and the two N-terminal
Ig domains of Robo, but the molecular details of this interaction and how it
induces signaling remain unclear. Here we describe the crystal structures of the
second LRR domain of human Slit2 (Slit2 D2), the first two Ig domains of its
receptor Robo1 (Ig1-2), and the minimal complex between these proteins (Slit2
D2-Robo1 Ig1). Slit2 D2 binds with its concave surface to the side of Ig1 with
electrostatic and hydrophobic contact regions mediated by residues that are
conserved in other family members. Surface plasmon resonance experiments and a
mutational analysis of the interface confirm that Ig1 is the primary domain for
binding Slit2. These structures provide molecular insight into Slit-Robo complex
formation and will be important for the development of novel cancer therapeutics.
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Figure 1.
Fig. 1. Structure of human Robo1 Ig1–2. (A) Ribbon
diagram. The disulfide bridges are in yellow, and the box
indicates the region highlighted in B. (B) Residues involved in
interdomain contacts of the Ig1-Ig2 interface. (C) Ribbon
diagram of the two Ig1–2 crystal forms showing the hinge
movement of Ig2. (D) Sequence alignment of Ig1 domains of human
Robo1, -2, -3, and -4 and of the Ig2 domain of human Robo1.
Residue numbering is for Robo1 Ig1 (above) and Robo1 Ig2
(below). Slit2 D2-binding residues selected for mutagenesis are
marked with a star, and residues strictly conserved between the
Ig1 domain of human Robo1, -2, -3, and -4 are shown in red.
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Figure 2.
Fig. 2. Structure of Slit2 D2 bound to Robo1 Ig1. Ig1 is in
green; Slit2 D2 N- and C-terminal caps are in purple and blue,
respectively; LRRs 1–6 are in orange; and the disulfide
bridges are in yellow. Interacting residues are shown in stick
representation. (A) Ribbon diagram of the complex in two
orthogonal orientations. (B) Electrostatic region of the Slit2
D2-Ig1 interface. (C) Hydrophobic region of the Slit2 D2-Ig1
interface. (D) Relative Slit2 D2-binding capacity of Robo1 Ig1
variants. Results are expressed as maximal Slit2 D2-binding,
normalized with respect to the maximal Slit2 D2-binding capacity
of wild-type Robo1 Ig1.
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Secondary reference #1
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Title
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Cloning, Expression, Crystallization and preliminary x-Ray analysis of the first two ig domains from human roundabout 1 (robo1).
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Authors
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C.Morlot,
W.Hemrika,
R.A.Romijn,
P.Gros,
S.Cusack,
A.A.Mccarthy.
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Ref.
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Acta Crystallogr Sect F Struct Biol Cryst Commun, 2007,
63,
689-691.
[DOI no: ]
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PubMed id
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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
percentage match of
96%.
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Figure 1.
Western blot and SDS --PAGE (15%) analysis of Robo1. (a)
Western blot analysis of media conditioned for 3 d revealed that
Robo1 Ig1 --2 is well expressed. (b) SDS --PAGE (15%) analysis
of purified Robo1 Ig1 --2. Acta Crystallogr Sect F Struct Biol
Cryst Commun. 2007 August 1; 63(Pt 8): 689–691. Published
online 2007 July 21. doi: 10.1107/S1744309107033027. Copyright
[copyright] International Union of Crystallography 2007
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Figure 2.
Plate-shaped crystals of Robo1 Ig1 --2 obtained in the
presence of KAuCl[4]. Acta Crystallogr Sect F Struct Biol Cryst
Commun. 2007 August 1; 63(Pt 8): 689–691. Published online
2007 July 21. doi: 10.1107/S1744309107033027. Copyright
[copyright] International Union of Crystallography 2007
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The above figures are
reproduced from the cited reference
which is an Open Access publication published by the IUCr
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