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Cellular and viral peptides bind multiple sites on the N-terminal domain of clathrin

DOI: 10.1111/tra.12457 DOI Help

Authors: Julia Muenzner (University of Cambridge) , Linton M. Traub (Department of Cell Biology, University of Pittsburgh School of Medicine) , Bernard T. Kelly (University of Cambridge) , Stephen Graham (University of Cambridge)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Traffic

State: Published (Approved)
Published: November 2016
Diamond Proposal Number(s): 8547 , 11235

Abstract: Short peptide motifs in unstructured regions of clathrin-adaptor proteins recruit clathrin to membranes to facilitate post-Golgi membrane transport. Three consensus clathrin-binding peptide sequences have been identified and structural studies show that each binds distinct sites on the clathrin heavy chain N-terminal domain (NTD). A fourth binding site for adaptors on NTD has been functionally identified but not structurally characterised. We have solved high resolution structures of NTD bound to peptide motifs from the cellular clathrin adaptors β2 adaptin and amphiphysin plus a putative viral clathrin adaptor, hepatitis D virus large antigen (HDAg-L). Surprisingly, with each peptide we observe simultaneous peptide binding at multiple sites on NTD and viral peptides binding to the same sites as cellular peptides. Peptides containing clathrin-box motifs (CBMs) with consensus sequence LΦxΦ[DE] bind at the ‘arrestin box’ on NTD, between β-propeller blades 4 and 5, which had previously been thought to bind a distinct consensus sequence. Further, we structurally define the fourth peptide binding site on NTD, which we term the Royle box. In vitro binding assays show that clathrin is more readily captured by cellular CBMs than by HDAg-L, and site-directed mutagenesis confirms that multiple binding sites on NTD contribute to efficient capture by CBM peptides.

Journal Keywords: Endocytosis; clathrin-mediated endocytosis; hepatitis D virus; amphiphysin; assembly polypeptide 2 (AP2); arrestin

Subject Areas: Biology and Bio-materials


Instruments: I02-Macromolecular Crystallography , I04-1-Macromolecular Crystallography (fixed wavelength)

Added On: 24/11/2016 13:45

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