PDBsum entry 3gc3

Endocytosis

PDB id: 3gc3

Contents

Protein chains

329 a.a. *

349 a.a. *

Waters ×276

* Residue conservation analysis

PDB id:

3gc3

Name:

Endocytosis

Title:

Crystal structure of arrestin2s and clathrin

Structure:

Beta-arrestin-1. Chain: a. Synonym: arrestin beta-1,arrestin-2. Engineered: yes. Clathrin heavy chain 1. Chain: b. Fragment: wd domain. Engineered: yes

Source:

Bos taurus. Cattle. Organism_taxid: 9913. Gene: arrb1. Expressed in: escherichia coli. Expression_system_taxid: 562. Gene: cltc.

Resolution:

2.20Å R-factor: 0.203 R-free: 0.251

Authors:

J.C.Williams,D.S.Kang

Key ref:

D.S.Kang et al. (2009). Structure of an arrestin2-clathrin complex reveals a novel clathrin binding domain that modulates receptor trafficking. J Biol Chem, 284, 29860-29872. PubMed id: 19710023 DOI: 10.1074/jbc.M109.023366

Date:

21-Feb-09 Release date: 25-Aug-09

Protein chain

P17870  (ARRB1_BOVIN) -  Beta-arrestin-1 from Bos taurus

Seq: 418 a.a.

Struc: 329 a.a.

Protein chain

P49951  (CLH1_BOVIN) -  Clathrin heavy chain 1 from Bos taurus

Seq: 1675 a.a.

Struc: 349 a.a.

Enzyme reactions

• Enzyme class: Chains A, B: E.C.?

DOI no: 10.1074/jbc.M109.023366

J Biol Chem 284:29860-29872 (2009)

PubMed id: 19710023

Structure of an arrestin2-clathrin complex reveals a novel clathrin binding domain that modulates receptor trafficking.

D.S.Kang, R.C.Kern, M.A.Puthenveedu, M.von Zastrow, J.C.Williams, J.L.Benovic.

ABSTRACT

Non-visual arrestins play a pivotal role as adaptor proteins in regulating the signaling and trafficking of multiple classes of receptors. Although arrestin interaction with clathrin, AP-2, and phosphoinositides contributes to receptor trafficking, little is known about the configuration and dynamics of these interactions. Here, we identify a novel interface between arrestin2 and clathrin through x-ray diffraction analysis. The intrinsically disordered clathrin binding box of arrestin2 interacts with a groove between blades 1 and 2 in the clathrin beta-propeller domain, whereas an 8-amino acid splice loop found solely in the long isoform of arrestin2 (arrestin2L) interacts with a binding pocket formed by blades 4 and 5 in clathrin. The apposition of the two binding sites in arrestin2L suggests that they are exclusive and may function in higher order macromolecular structures. Biochemical analysis demonstrates direct binding of clathrin to the splice loop in arrestin2L, whereas functional analysis reveals that both binding domains contribute to the receptor-dependent redistribution of arrestin2L to clathrin-coated pits. Mutagenesis studies reveal that the clathrin binding motif in the splice loop is (L/I)(2)GXL. Taken together, these data provide a framework for understanding the dynamic interactions between arrestin2 and clathrin and reveal an essential role for this interaction in arrestin-mediated endocytosis.

Selected figure(s)

Figure 4.
Structure of an arrestin2L-(1–393) complex with clathrin-(1–363).A, ribbon model of the complex shows a molecule of clathrin-(1–363) (green) with two molecules of arrestin2L-(1–393) (cyan) using two independent binding interfaces. The boxed area (red) is an additional interface using the splice loop of arrestin2L. B, multiwavelength anomalous diffraction electron density maps (σ = 1.5) showing the splice loop of arrestin2L (top) and detailed structure of the boxed area from A with labeled key residues of arrestin2L and clathrin (bottom; black and green, respectively). C, B is depicted as a one-dimensional map noting possible hydrogen bonding (dotted lines) and hydrophobic interactions (bold dotted lines).

Figure 7.
Comparison of arrestin position in clathrin superposition.A, top view of the clathrin terminal domain shows two distinct interactions with arrestin2L, which are depicted as red ribbons (left). The side view of arrestin2L shows two separated clathrin binding sites, the LϕXϕ(D/E) motif and splice loop (right, red dotted lines). B, side view; arrestins (cyan, low resolution; wheat, high resolution) near the clathrin binding box (red ribbon) do not superimpose upon the superposition of clathrin (green) from the different crystal forms. Also shown is the second arrestin with a splice loop. Each end point of flanking regions in unstructured C-terminal loop is marked and connected as blue balls and a dotted line for the N-terminal area and red balls and a dotted line for the C-terminal area. C, top view; same as B but rotated 90° in a plane and looking down at approximate rotation axis relating the two arrestins. D, electron microscopy image of a clathrin barrel with a set of clathrin terminal domains shown in red (from Edeling et al. (59)). The enlarged inset is a schematic representation of the six terminal domains within the cluster, depicting a distance of ∼64 Å between terminal domains. The distance between terminal domains was measured using the measurement function in PyMol using PDB code 1XI4 for the clathrin D6 coat (48).

The above figures are reprinted by permission from the ASBMB: J Biol Chem (2009, 284, 29860-29872) copyright 2009.

Figures were selected by the author.