PDBsum entry 2v8f

Protein binding

PDB id: 2v8f

Contents

Protein chains

139 a.a. *

16 a.a. *

Ligands

SO4 ×5

GOL

IPA

Metals

_NA

Waters ×357

* Residue conservation analysis

PDB id:

2v8f

Name:

Protein binding

Title:

Mouse profilin iia in complex with a double repeat from the fh1 domain of mdia1

Structure:

Profilin-2. Chain: a, b. Synonym: profilin iia. Engineered: yes. Protein diaphanous homolog 1. Chain: c. Fragment: fh1 domain, residues 635-655. Synonym: diaphanous-related formin-1, drf1, p140mdia, mdia1. Engineered: yes.

Source:

Mus musculus. House mouse. Organism_taxid: 10090. Expressed in: escherichia coli. Expression_system_taxid: 562. Synthetic: yes. Organism_taxid: 10090

Resolution:

1.10Å R-factor: 0.146 R-free: 0.173

Authors:

P.Kursula,I.Kursula,J.Downer,W.Witke,M.Wilmanns

Key ref:

P.Kursula et al. (2008). High-resolution structural analysis of mammalian profilin 2a complex formation with two physiological ligands: the formin homology 1 domain of mDia1 and the proline-rich domain of VASP. J Mol Biol, 375, 270-290. PubMed id: 18001770 DOI: 10.1016/j.jmb.2007.10.050

Date:

07-Aug-07 Release date: 18-Dec-07

Protein chains

Q9JJV2  (PROF2_MOUSE) -  Profilin-2 from Mus musculus

Seq: 140 a.a.

Struc: 139 a.a.*

Protein chain

O08808  (DIAP1_MOUSE) -  Protein diaphanous homolog 1 from Mus musculus

Seq: 1255 a.a.

Struc: 16 a.a.

* PDB and UniProt seqs differ at 1 residue position (black cross)

DOI no: 10.1016/j.jmb.2007.10.050

J Mol Biol 375:270-290 (2008)

PubMed id: 18001770

High-resolution structural analysis of mammalian profilin 2a complex formation with two physiological ligands: the formin homology 1 domain of mDia1 and the proline-rich domain of VASP.

P.Kursula, I.Kursula, M.Massimi, Y.H.Song, J.Downer, W.A.Stanley, W.Witke, M.Wilmanns.

ABSTRACT

Profilins are small proteins capable of binding actin, poly-l-proline and other proline-rich sequences, and phosphatidylinositol (4,5)-bisphosphate. A number of proline-rich ligands for profilin have been characterised, including proteins of the Ena/VASP and formin families. We have determined the high-resolution crystal structures of mouse profilin 2a in complex with peptides from two functionally important ligands from different families, VASP and mDia1. The structures show that the binding mode of the peptide ligand is strongly affected by the non-proline residues in the sequence, and the peptides from VASP and mDia1 bind to profilin 2a in distinct modes. The high resolution of the crystallographic data allowed us to detect conserved CH-pi hydrogen bonds between the peptide and profilin in both complexes. Furthermore, both peptides, which are shown to have micromolar affinity, induced the dimerisation of profilin, potentially leading to functionally different ligand-profilin-actin complexes. The peptides did not significantly affect actin polymerisation kinetics in the presence or in the absence of profilin 2a. Mutant profilins were tested for binding to poly-L-proline and the VASP and mDia1 peptides, and the F139A mutant bound proline-rich ligands with near-native affinity. Peptide blotting using a series of designed peptides with profilins 1 and 2a indicates differences between the two profilins towards proline-rich peptides from mDia1 and VASP. Our data provide structural insights into the mechanisms of mDia1 and VASP regulated actin polymerisation.

Selected figure(s)

Figure 1.
Figure 1. The profilin ligands used in the current study. (a) A diagram of the structures of the profilin 2a ligands, mDia1 and VASP. For mDia1, the domains are as follows: GBD, Rho-GTPase-binding domain; DID, diaphanous inhibitory domain; DD, dimerisation domain; CC, coiled-coil domain; FH1, formin homology domain 1; FH2, formin homology domain 2; DAD, diaphanous autoinhibitory domain. The regions N-terminal to the FH1 domain have been referred to in the literature as the FH3 domain. For VASP, the EVH1 domain, the proline-rich domain, and the EVH2 domain are indicated. The EVH2 domain contains a WH2 domain (dark grey), an F-actin binding domain (light grey), and a tetramerisation domain (white). The sequences of the peptides used in this study are shown below the profilin-binding domains. (b) The sequence of the FH1 domain of mDia1, which has 13 highly homologous proline-rich repeats arranged in tandem. The peptide used in this study, containing two binding motifs, is underlined.

Figure 3.
Figure 3. Binding site determinants. (a) Stereo view of the binding site for the VASP peptide, showing hydrogen bond interactions and several residues discussed in the text. (b) Superposition of the peptide binding modes from the VASP (proline, orange; glycine, red) and mDia1 (proline, green; non-proline, blue) complexes. The N termini of the peptides point right in the Figure.

The above figures are reprinted by permission from Elsevier: J Mol Biol (2008, 375, 270-290) copyright 2008.

Figures were selected by an automated process.