PDBsum entry 2v9t

Structural protein/receptor

PDB id

2v9t

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Contents

			Protein chains

					107 a.a. *


					210 a.a. *

			Waters ×385

* Residue conservation analysis

PDB id:

2v9t

Links

Name:

Structural protein/receptor

Title:

Complex between the second lrr domain of slit2 and the first ig domain from robo1

Structure:

Roundabout homolog 1. Chain: a. Fragment: ig1-2m, residues 61-166. Synonym: h-robo-1, deleted in u twenty twenty, robo1. Engineered: yes. Mutation: yes. Slit homolog 2 protein n-product. Chain: b. Fragment: second lrr domain, residues 271-479.

Source:

Homo sapiens. Human. Organism_taxid: 9606. Expressed in: homo sapiens. Expression_system_taxid: 9606. Expression_system_cell_line: hek293-ebna1.

Resolution:

1.70Å

R-factor:

0.188

R-free:

0.226

Authors:

C.Morlot,S.Cusack,A.A.Mccarthy

Key ref:

C.Morlot et al. (2007). Structural insights into the Slit-Robo complex. Proc Natl Acad Sci U S A, 104, 14923-14928. PubMed id: 17848514 DOI: 10.1073/pnas.0705310104

Date:

25-Aug-07

Release date:

25-Sep-07

PROCHECK

	Headers

	References

Protein chain

Q9Y6N7 (ROBO1_HUMAN) - Roundabout homolog 1 from Homo sapiens

Seq: Struc:

Seq: Struc:

Seq: Struc:

Seq: Struc:					1651 a.a. 107 a.a.*

Protein chain

O94813 (SLIT2_HUMAN) - Slit homolog 2 protein from Homo sapiens

Seq: Struc:

Seq: Struc:

Seq: Struc:

Seq: Struc:					1529 a.a. 210 a.a.*

Key:

PfamA domain

Secondary structure

CATH domain

* PDB and UniProt seqs differ at 4 residue positions (black crosses)

DOI no: 10.1073/pnas.0705310104	Proc Natl Acad Sci U S A 104:14923-14928 (2007)
PubMed id: 17848514

Structural insights into the Slit-Robo complex.
C.Morlot, N.M.Thielens, R.B.Ravelli, W.Hemrika, R.A.Romijn, P.Gros, S.Cusack, A.A.McCarthy.

ABSTRACT

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.

Selected figure(s)



	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.		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.

Figures were selected by an automated process.

Literature references that cite this PDB file's key reference

PubMed id

Reference

21402080

B.H.Biersmith, M.Hammel, E.R.Geisbrecht, and S.Bouyain (2011).
The immunoglobulin-like domains 1 and 2 of the protein tyrosine phosphatase LAR adopt an unusual horseshoe-like conformation.

J Mol Biol, 408, 616-627.

PDB codes:

3pxh 3pxj

20298552

A.E.Denk, S.Kaufmann, K.Stark, J.Schedel, T.Lowin, T.Schubert, and A.K.Bosserhoff (2010).
Slit3 inhibits Robo3-induced invasion of synovial fibroblasts in rheumatoid arthritis.

Arthritis Res Ther, 12, R45.

20452960

R.H.Adams, and A.Eichmann (2010).
Axon guidance molecules in vascular patterning.

Cold Spring Harb Perspect Biol, 2, a001875.

19934013

C.E.Shiau, and M.Bronner-Fraser (2009).
N-cadherin acts in concert with Slit1-Robo2 signaling in regulating aggregation of placode-derived cranial sensory neurons.

Development, 136, 4155-4164.

19498462

E.Seiradake, A.C.von Philipsborn, M.Henry, M.Fritz, H.Lortat-Jacob, M.Jamin, W.Hemrika, M.Bastmeyer, S.Cusack, and A.A.McCarthy (2009).
Structure and functional relevance of the Slit2 homodimerization domain.

EMBO Rep, 10, 736-741.

PDB code:

2wfh

19608877

F.Shen, X.Liu, J.G.Geng, and S.W.Guo (2009).
Increased immunoreactivity to SLIT/ROBO1 in ovarian endometriomas: a likely constituent biomarker for recurrence.

Am J Pathol, 175, 479-488.

19452560

K.L.Hindle, J.Bella, and S.C.Lovell (2009).
Quantitative analysis and prediction of curvature in leucine-rich repeat proteins.

Proteins, 77, 342-358.

19350278

S.Zu, Z.Bartik, S.Zhao, U.Sillen, and A.Nordenskjöld (2009).
Mutations in the ROBO2 and SLIT2 genes are rare causes of familial vesico-ureteral reflux.

Pediatr Nephrol, 24, 1501-1508.

18345009

C.A.Jones, N.R.London, H.Chen, K.W.Park, D.Sauvaget, R.A.Stockton, J.D.Wythe, W.Suh, F.Larrieu-Lahargue, Y.S.Mukouyama, P.Lindblom, P.Seth, A.Frias, N.Nishiya, M.H.Ginsberg, H.Gerhardt, K.Zhang, and D.Y.Li (2008).
Robo4 stabilizes the vascular network by inhibiting pathologic angiogenesis and endothelial hyperpermeability.

Nat Med, 14, 448-453.

18264786

J.A.Legg, J.M.Herbert, P.Clissold, and R.Bicknell (2008).
Slits and Roundabouts in cancer, tumour angiogenesis and endothelial cell migration.

Angiogenesis, 11, 13-21.

18359766

N.Fukuhara, J.A.Howitt, S.A.Hussain, and E.Hohenester (2008).
Structural and functional analysis of slit and heparin binding to immunoglobulin-like domains 1 and 2 of Drosophila Robo.

J Biol Chem, 283, 16226-16234.

PDB codes:

2vr9 2vra

18214954

Q.R.Fan, and W.A.Hendrickson (2008).
Comparative structural analysis of the binding domain of follicle stimulating hormone receptor.

Proteins, 72, 393-401.

18980679

S.Kaur, G.V.Samant, K.Pramanik, P.W.Loscombe, M.L.Pendrak, D.D.Roberts, and R.Ramchandran (2008).
Silencing of directional migration in roundabout4 knockdown endothelial cells.

BMC Cell Biol, 9, 61.

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.