TY - JOUR
T1 - Location of auxilin within a clathrin cage
AU - Smith, CJ
AU - Dafforn, Timothy
AU - Kent, H
AU - Sims, CA
AU - Khubchandani-Aswani, K
AU - Zhang, L
AU - Saibil, HR
AU - Pearse, BM
PY - 2004/2/13
Y1 - 2004/2/13
N2 - The Dna J homologue, auxilin, acts as a co-chaperone for Hsc70 in the uncoating of clathrin-coated vesicles during endocytosis. Biochemical studies have aided understanding of the uncoating mechanism but until now there was no structural information on how auxilin interacts with the clathrin cage. Here we have determined the three-dimensional structure of a complex of auxilin with clathrin cages by cryo-electron microscopy and single particle analysis. We show that auxilin forms a discrete shell of density on the inside of the clathrin cage. Peptide competition assays confirm that a candidate clathrin box motif in auxilin, LLGLE, can bind to a clathrin construct containing the beta-propeller domain and also displace the well-characterised LLNLD clathrin box motif derived from the beta-adaptin hinge region. The means by which auxilin could both aid clathrin coat assembly and displace clathrin from AP2 during uncoating is discussed.
AB - The Dna J homologue, auxilin, acts as a co-chaperone for Hsc70 in the uncoating of clathrin-coated vesicles during endocytosis. Biochemical studies have aided understanding of the uncoating mechanism but until now there was no structural information on how auxilin interacts with the clathrin cage. Here we have determined the three-dimensional structure of a complex of auxilin with clathrin cages by cryo-electron microscopy and single particle analysis. We show that auxilin forms a discrete shell of density on the inside of the clathrin cage. Peptide competition assays confirm that a candidate clathrin box motif in auxilin, LLGLE, can bind to a clathrin construct containing the beta-propeller domain and also displace the well-characterised LLNLD clathrin box motif derived from the beta-adaptin hinge region. The means by which auxilin could both aid clathrin coat assembly and displace clathrin from AP2 during uncoating is discussed.
UR - http://www.scopus.com/inward/record.url?scp=0742306907&partnerID=8YFLogxK
U2 - 10.1016/j.jmb.2003.12.006
DO - 10.1016/j.jmb.2003.12.006
M3 - Article
C2 - 14757058
VL - 336
SP - 461
EP - 471
JO - Journal of Molecular Biology
JF - Journal of Molecular Biology
ER -