Molecular Architecture and Functional Model of the Endocytic AP2 Complex

Collins, Brett M.; McCoy, Airlie J.; Kent, Helen M.; Evans, Philip R.; Owen, David J.

doi:10.1016/s0092-8674(02)00735-3

Cited by 558 publications

(684 citation statements)

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“…The molecular details underlying these apparent differences between mammalian AP2 and yeast AP3 require detailed structural information on AP3. It is noteworthy that the chain and the N-terminal region of the mu chain of adaptins contain a longin fold (Collins et al, 2002;Heldwein et al, 2004) as apparently do the and ␦ subunits of the COPI coat protein complex (Schlenker et al, 2006). The significance of a shared fold in the vesicle coats and cargo sorting complexes that presumably recognize and sort longin SNAREs merits investigation.…”

Section: Discussionmentioning

confidence: 99%

“…The longin domain (LD) is not restricted to R-SNAREs and has been identified in a variety of proteins (Rossi et al, 2004), some of which are also involved in vesicular transport. These include subunits of the adaptin complexes (Collins et al, 2002;Heldwein et al, 2004) and SEDL/Trs20p (Jang et al, 2002), a common subunit of the transport protein particle (TRAPP)I and TRAPPII multisubunit complexes, which are required for traffic between the ER and Golgi and within the Golgi (Oka and Krieger, 2005). The function of the longin fold of Sec22b/Sec22p is unknown, but in Ykt6p the longin domain folds back and binds to the SNARE-motif of the protein (Tochio et al, 2001), and this conformation is likely to be important for chaperoning the lipid modified C terminus of the cytoplasmic form of Ykt6p in cells as well as for its localization (Fukasawa et al, 2004;Hasegawa et al, 2004).…”

Section: Introductionmentioning

confidence: 99%

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Identification of the Yeast R-SNARE Nyv1p as a Novel Longin Domain-containing Protein

Wen¹,

Chen

Wu³

et al. 2006

MBoC

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Identification of the Yeast R-SNARE Nyv1p as a Novel Longin Domain-containing Protein

Wen¹,

Chen

Wu³

et al. 2006

MBoC

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“…The null mutant of the m-subunit still accumulates the a-and s-subunits, and the double mutant lacking m-and a-subunits is embryo lethal (Gu et al, 2013). It has been suggested that the a/s-and b/m-subunits have the potential to form large/small subunit heterodimers (Collins et al, 2002;Jackson et al, 2010), which may partially compensate for the loss of AP-2 (Gu et al, 2013). Such mechanisms might complement the lack of AP-2 function in the ap2m mutant plants.…”

Section: Homozygous Ap2m Mutant Plants Are Viablementioning

confidence: 99%

Identification and Dynamics of Arabidopsis Adaptor Protein-2 Complex and Its Involvement in Floral Organ Development

et al. 2013

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ORCID ID: 0000-0003-4154-9967 (S.Y.).The adaptor protein-2 (AP-2) complex is a heterotetramer involved in clathrin-mediated endocytosis of cargo proteins from the plasma membrane in animal cells. The homologous genes of AP-2 subunits are present in the genomes of plants; however, their identities and roles in endocytic pathways are not clearly defined in plants. Here, we reveal the molecular composition of the AP-2 complex of Arabidopsis thaliana and its dynamics on the plasma membrane. We identified all of the a-, b-, s-, and m-subunits of the AP-2 complex and detected a weak interaction of the AP-2 complex with clathrin heavy chain. The m-subunit protein fused to green fluorescent protein (AP2M-GFP) was localized to the plasma membrane and to the cytoplasm. Live-cell imaging using a variable-angle epifluorescence microscope revealed that AP2M-GFP transiently forms punctate structures on the plasma membrane. Homozygous ap2m mutant plants exhibited abnormal floral structures, including reduced stamen elongation and delayed anther dehiscence, which led to a failure of pollination and a subsequent reduction of fertility. Our study provides a molecular basis for understanding AP-2-dependent endocytic pathways in plants and their roles in floral organ development and plant reproduction.

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“…Vps35 resembles many other helical solenoid proteins, including other important players in coated vesicle trafficking. These include the trunk domains of the large subunits of the AP adaptor protein complexes 18,19 (Fig. S3), whose function is to link clathrin to cargo and membranes, the helical repeat regions of the heavy chain of clathrin itself 20,21, and the Sec13/Sec31 assembly unit of the COPII coat 22.…”

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confidence: 99%

Functional architecture of the retromer cargo-recognition complex

Hierro

Rojas

et al. 2007

Nature

265

381

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The retromer complex 1,2 is required for the sorting of acid hydrolases to lysosomes 3-7, transcytosis of the polymeric Ig receptor 8, Wnt gradient formation 9,10, iron transporter recycling 11, and processing of the amyloid precursor protein 12. Human retromer consists of two smaller complexes, the cargo recognition Vps26:Vps29:Vps35 heterotrimer, and a membrane-targeting heterodimer or homodimer of SNX1 and/or SNX2 13. The crystal structure of a Vps29:Vps35 subcomplex shows how the metallophosphoesterase-fold subunit Vps29 14,15 acts as a scaffold for the C-terminal half of Vps35. Vps35 forms a horseshoe-shaped right-handed α-helical solenoid whose concave face completely covers the metal-binding site of Vps29 and whose convex face exposes a series of hydrophobic interhelical grooves. Electron microscopy shows that the intact Vps26:Vps29:Vps35 complex is a stick-shaped, somewhat flexible, structure, ∼ 21 nm long. A hybrid structural model derived from crystal structures, electron microscopy, interaction studies, and bioinformatics shows that the α-solenoid fold extends the full length of Vps35, and that Vps26 is bound at the opposite end from Vps29. This extended structure presents multiple binding sites for the SNX complex and receptor cargo, and appears capable of flexing to conform to curved vesicular membranes.The retromer cargo recognition complex consists of the 38-kDa Vps26, 20-kDa Vps29, and 92-kDa Vps35 subunits. The structures of the two smaller subunits have been determined in isolation. Vps26 is a structural cousin of the arrestins 16, a family of trafficking proteins that directly bind to cell surface receptors and direct their internalization. Vps29 has a metallophosphoesterase fold 14,15 that can bind two metal ions. Compared to functional metallophosphoesterases, a key His that serves as a catalytic base in the 6To whom correspondence should be addressed:James H. Hurley, (301) 402−4703, fax (301) 480−0639; E-mail: hurley@helix.nih.gov. 2 These authors contributed equally. Author contributions A. L. R. and A. H. expressed and purified protein complexes, crystallized the Vps29:Vps35 C-terminal subcomplex, collected crystallographic data, and determined and refined the crystal structure; A. H. carried out phosphatase assays; R. R. and N. M. carried out immunoprecipitation and optical microscopy studies; G. E. and A. C. S. carried out and interpreted electron microscopy studies; A. V. K. carried out sequence analysis; and J. H. H., J. S. B., and A. C. S. designed the study. A. H. and A. L. R. contributed equally to this study. Here we take a structural approach to gain insight into the function of retromer. Author informationThe crystal structure of a Vps29:Vps35 subcomplex, containing the C-terminal 40% of the large Vps35 subunit, was determined at 2.8 Å resolution (Fig. 1a, Fig. S1, Table S1). The Cterminal portion of Vps35 consists of a single right-handed superhelix with a pitch of 12 Å and a total of 13 helices (Fig. 1a, S2). Vps35 resembles many other helical solenoid proteins, includ...

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Molecular Architecture and Functional Model of the Endocytic AP2 Complex

Cited by 558 publications

References 59 publications

Identification of the Yeast R-SNARE Nyv1p as a Novel Longin Domain-containing Protein

Identification of the Yeast R-SNARE Nyv1p as a Novel Longin Domain-containing Protein

Identification and Dynamics of Arabidopsis Adaptor Protein-2 Complex and Its Involvement in Floral Organ Development

Functional architecture of the retromer cargo-recognition complex

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