PDBsum entry 5jm6

Hydrolase

PDB id: 5jm6

Contents

Protein chains

(+ 0 more) 445 a.a.

Metals

_ZN ×12

Waters ×279

PDB id:

5jm6

Name:

Hydrolase

Title:

Structure of chaetomium thermophilum mape1

Structure:

Aminopeptidase-like protein. Chain: a, e, f, b, c, d. Engineered: yes

Source:

Chaetomium thermophilum. Organism_taxid: 209285. Gene: ctht_0065300. Expressed in: escherichia coli bl21(de3). Expression_system_taxid: 469008. Expression_system_variant: ril.

Resolution:

2.76Å R-factor: 0.209 R-free: 0.242

Authors:

C.Bertipaglia,A.J.Jakobi,M.Wilmanns,C.Sachse

Key ref:

C.Bertipaglia et al. (2016). Higher-order assemblies of oligomeric cargo receptor complexes form the membrane scaffold of the Cvt vesicle. Embo Rep, 17, 1044-1060. PubMed id: 27266708 DOI: 10.15252/embr.201541960

Date:

28-Apr-16 Release date: 15-Jun-16

Protein chains

G0SG74  (G0SG74_CHATD) -  Aminopeptidase-like protein from Chaetomium thermophilum (strain DSM 1495 / CBS 144.50 / IMI 039719)

Seq: 519 a.a.

Struc: 445 a.a.

DOI no: 10.15252/embr.201541960

Embo Rep 17:1044-1060 (2016)

PubMed id: 27266708

Higher-order assemblies of oligomeric cargo receptor complexes form the membrane scaffold of the Cvt vesicle.

C.Bertipaglia, S.Schneider, A.J.Jakobi, A.K.Tarafder, Y.S.Bykov, A.Picco, W.Kukulski, J.Kosinski, W.J.Hagen, A.C.Ravichandran, M.Wilmanns, M.Kaksonen, J.A.Briggs, C.Sachse.

ABSTRACT

Selective autophagy is the mechanism by which large cargos are specifically sequestered for degradation. The structural details of cargo and receptor assembly giving rise to autophagic vesicles remain to be elucidated. We utilize the yeast cytoplasm-to-vacuole targeting (Cvt) pathway, a prototype of selective autophagy, together with a multi-scale analysis approach to study the molecular structure of Cvt vesicles. We report the oligomeric nature of the major Cvt cargo Ape1 with a combined 2.8 Å X-ray and negative stain EM structure, as well as the secondary cargo Ams1 with a 6.3 Å cryo-EM structure. We show that the major dodecameric cargo prApe1 exhibits a tendency to form higher-order chain structures that are broken upon interaction with the receptor Atg19 in vitro The stoichiometry of these cargo-receptor complexes is key to maintaining the size of the Cvt aggregate in vivo Using correlative light and electron microscopy, we further visualize key stages of Cvt vesicle biogenesis. Our findings suggest that Atg19 interaction limits Ape1 aggregate size while serving as a vehicle for vacuolar delivery of tetrameric Ams1.