PDBsum entry 1bb9

Transferase

PDB id: 1bb9

Contents

Protein chain

83 a.a. *

Waters ×60

* Residue conservation analysis

PDB id:

1bb9

Name:

Transferase

Title:

Crystal structure of the sh3 domain from rat amphiphysin 2

Structure:

Amphiphysin 2. Chain: a. Fragment: sh3 domain. Engineered: yes

Source:

Rattus norvegicus. Norway rat. Organism_taxid: 10116. Organ: brain. Expressed in: escherichia coli bl21. Expression_system_taxid: 511693.

Resolution:

2.20Å R-factor: 0.186 R-free: 0.264

Authors:

D.J.Owen,H.T.Mcmahon,P.R.Evans

Key ref:

D.J.Owen et al. (1998). Crystal structure of the amphiphysin-2 SH3 domain and its role in the prevention of dynamin ring formation. Embo J, 17, 5273-5285. PubMed id: 9736607 DOI: 10.1093/emboj/17.18.5273

Date:

29-Apr-98 Release date: 17-Jun-98

Protein chain

O08839  (BIN1_RAT) -  Myc box-dependent-interacting protein 1 from Rattus norvegicus

Seq: 588 a.a.

Struc: 83 a.a.

Enzyme reactions

• Enzyme class: E.C.?

DOI no: 10.1093/emboj/17.18.5273

Embo J 17:5273-5285 (1998)

PubMed id: 9736607

Crystal structure of the amphiphysin-2 SH3 domain and its role in the prevention of dynamin ring formation.

D.J.Owen, P.Wigge, Y.Vallis, J.D.Moore, P.R.Evans, H.T.McMahon.

ABSTRACT

The amphiphysins are brain-enriched proteins, implicated in clathrin-mediated endocytosis, that interact with dynamin through their SH3 domains. To elucidate the nature of this interaction, we have solved the crystal structure of the amphiphysin-2 (Amph2) SH3 domain to 2.2 A. The structure possesses several notable features, including an extensive patch of negative electrostatic potential covering a large portion of its dynamin binding site. This patch accounts for the specific requirement of amphiphysin for two arginines in the proline-rich binding motif to which it binds on dynamin. We demonstrate that the interaction of dynamin with amphiphysin SH3 domains, unlike that with SH3 domains of Grb2 or spectrin, prevents dynamin self-assembly into rings. Deletion of a unique insert in the n-Src loop of Amph2 SH3, a loop adjacent to the dynamin binding site, significantly reduces this effect. Conversely, replacing the n-Src loop of the N-terminal SH3 domain of Grb2 with that of Amph2 causes it to favour dynamin ring disassembly. Transferrin uptake assays show that shortening the n-Src loop of Amph2 SH3 reduces the ability of this domain to inhibit endocytosis in vivo. Our data suggest that amphiphysin SH3 domains are important regulators of the multimerization cycle of dynamin in endocytosis.

Selected figure(s)

Figure 4.
Figure 4 Surface representations of SH3 domains from Abl, Sem5 and Amph2. The pictures on the left depict surface accessible hydrophobic regions coloured green to yellow for increasing hydrophobicity (M.Noble, X objects, unpublished). Amph2 SH3 shows only two hydrophobic patches as opposed to the three present in Abl and Sem5. On the right are representations of electrostatic potential (created using GRASP) showing the large negatively charged patch (red) on the peptide binding surface (the peptide is shown in the case of Abl and Sem5). The representations show the peptide binding surfaces of the SH3 domains with the n-Src loops pointing toward the top of the page. Coordinates for the SH3 domain of Abl (Musacchio et al., 1994) and the N-terminal SH3 domain of sem5 (Lim et al., 1994) were obtained from the Protein Data Bank.

Figure 5.
Figure 5 Point mutations in the Amph2 SH3 domain. (A) Structure of Amph2 SH3 showing positions of key mutated residues, coloured according to type. Hydrophobic residues are coloured green and acidic residues in magenta. The longer n-Src loop which is unique to the amphiphysins (the DAPS) that is exchanged for the shorter homologous loop from Grb2 NSH3 is coloured blue. (B) Effect of point mutations in GST Amph2 SH3 domain on its ability to bind dynamin. (C) Interaction between Amph2 SH3 and dynamin is sensitive to pH. Binding to wild-type is maximal at pH 7.0, but rapidly declines as pH is either increased or decreased.

The above figures are reprinted from an Open Access publication published by Macmillan Publishers Ltd: Embo J (1998, 17, 5273-5285) copyright 1998.

Figures were selected by an automated process.