A Third Specificity‐Determining Site in μ2 Adaptin for Sequences Upstream of YxxΦ Sorting Motifs

Owen, David J.; Setiadi, Hendra; Evans, Philip R.; McEver, Rodger P.; Green, Samuel A.

doi:10.1034/j.1600-0854.2001.020205.x

Cited by 47 publications

(43 citation statements)

References 38 publications

(61 reference statements)

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“…In the absence of a bulky hydrophobic residue at Yϩ3, amino acids with bulky hydrophobic side chains at YϪ3 have also been reported to be important for 2 binding (26). However, we found that a valine residue at YϪ3 in P2X 4 was not required for endocytosis.…”

Section: Discussioncontrasting

confidence: 50%

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Identification of a Non-canonical Tyrosine-based Endocytic Motif in an Ionotropic Receptor

Royle

Bobanović

Murrell‐Lagnado

2002

Journal of Biological Chemistry

100

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Rapid modulation of the surface number of certain ionotropic receptors is achieved by altering the relative rates of insertion and internalization. These receptors are internalized by a clathrin-mediated pathway; however, a motif that is necessary for endocytosis of ionotropic receptors has not yet been identified. Here, we identified a motif that is required for constitutive and agonist-regulated internalization of the ionotropic P2X 4 receptor. Three amino acids in the C terminus of P2X 4 (Tyr 378 , Gly 381 , and Leu 382 ) compose a non-canonical tyrosine-based sorting signal of the form YXXGL. We found that P2X 4 protein was present in clathrin-coated vesicles isolated from rat brain and that a glutathione S-transferase fusion of the P2X 4 C terminus pulled down the adaptor protein-2 complex from brain extract. Mutation of either the tyrosine-binding pocket of the 2 subunit of adaptor protein-2 or the YXXGL motif in the receptor C terminus caused a decrease in receptor internalization and a dramatic increase in the surface expression of P2X 4 receptors. The YXXGL motif represents a non-canonical tyrosine-based sorting signal that is necessary for efficient endocytosis of the P2X 4 receptor. Similar motifs are present in other receptors and may be important for the control of their functional expression.

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Section: Discussioncontrasting

confidence: 50%

“…We propose that, like YXXØ motifs, Tyr 378 and Leu 382 of P2X 4 fit into the two hydrophobic binding pockets on either side of strand ␤16 (12,25,26). If the C terminus of P2X 4 were in an extended conformation, this fit would not be favorable.…”

Section: Discussionmentioning

confidence: 99%

Identification of a Non-canonical Tyrosine-based Endocytic Motif in an Ionotropic Receptor

Royle

Bobanović

Murrell‐Lagnado

2002

Journal of Biological Chemistry

100

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“…In addition to the NPxY/NxxY SNX17-binding signal, the ICD of Pselectin also contains an overlapping Yxxϕ motif ( 755 YGVF 758 in P-selectin) recognized by the activating protein 2 (AP2) clathrin adaptor for endocytosis, presenting an interesting example of how overlapping sorting sequences can be used by different trafficking machineries. The structure of the AP2 μ2 subunit in complex with the P-selectin YGVF peptide has been determined by X-ray crystallography (38), allowing a comparison with the SNX17-bound P-selectin ICD containing both trafficking signals (GTYGVFTNAAYDPT) to investigate any similarities in the peptide-binding mechanisms (Fig. 3C).…”

Section: Resultsmentioning

confidence: 99%

“…(C) The Pselectin ICD contains overlapping Yxxϕ and NPxY/NxxY sorting motifs. The structure of the P-selectin ICD bound to SNX17 (yellow) is overlaid with the contiguous P-selectin YGVF-containing peptide bound to the AP2 μ2 subunit (magenta) (38).…”

Section: Px-ferm Proteins Bind Promiscuously To a Large Variety Of Pumentioning

confidence: 99%

Structural basis for endosomal trafficking of diverse transmembrane cargos by PX-FERM proteins

Ghai

Bugarčić

Liu

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

119

142

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Transit of proteins through the endosomal organelle following endocytosis is critical for regulating the homeostasis of cell-surface proteins and controlling signal transduction pathways. However, the mechanisms that control these membrane-transport processes are poorly understood. The Phox-homology (PX) domain-containing proteins sorting nexin (SNX) 17, SNX27, and SNX31 have emerged recently as key regulators of endosomal recycling and bind conserved Asn-Pro-Xaa-Tyr-sorting signals in transmembrane cargos via an atypical band, 4.1/ezrin/radixin/moesin (FERM) domain. Here we present the crystal structure of the SNX17 FERM domain bound to the sorting motif of the P-selectin adhesion protein, revealing both the architecture of the atypical FERM domain and the molecular basis for recognition of these essential sorting sequences. We further show that the PX-FERM proteins share a promiscuous ability to bind a wide array of putative cargo molecules, including receptor tyrosine kinases, and propose a model for their coordinated molecular interactions with membrane, cargo, and regulatory proteins.endosome | protein crystallography | X-ray scattering | membrane trafficking T he cell-surface levels of signaling and adhesion receptors, nutrient transporters, ion channels, and many other proteins are tightly regulated by opposing endocytic, exocytic, and endosomal recycling transport pathways. The selective sorting of these transmembrane proteins is the consequence of their interaction with essential adaptor proteins via conserved motifs present in their cytosolic tails, generally based on short linear amino acid sequences such as the Yxxϕ, DxxLL, and [DE]xxxL [LI] motifs (where ϕ is any bulky hydrophobic side-chain, and x is any residue) recognized by clathrin adaptors (1-3). The first identified sorting signal was the Asn-Pro-Xaa-Tyr (NPxY) motif, initially described in the LDL receptor (LDLR) isolated from patients suffering from familial hypercholesterolemia, where mutation of the sequence results in defective internalization and cholesterol uptake (4). The recent structure of the autosomal recessive hypercholesterolemia protein (ARH) phosphotyrosinebinding (PTB) domain in complex with the LDLR intracellular domain (ICD) provides the molecular basis of LDLR recognition and internalization within clathrin-coated pits by endocytic adaptor proteins (5).Although little is known about the sorting sequences and mechanisms required for competing endosome-to-cell surface recycling pathways, emerging evidence shows that the NPxY motif not only is vital to internalization but also aids cargo recycling via organelle-specific recognition by the endosomal protein, sorting nexin 17 (SNX17). SNX17 is a member of the Phox-homology (PX) domain-containing protein family and has been shown to be a critical regulator of endosomal sorting and cell-surface recycling of several essential cargo molecules. These include P-selectin (6-8), amyloid precursor protein (APP) (9, 10), integrins (11,12), and members of the LDL receptor family (13-17) ...

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“…Both large adaptins bind clathrin and recruit additional socalled ''accessory'' proteins to the site of transport vesicle formation. Besides m1, cargo proteins are also bound by b1, g1/ s1 hemi-complexes and s1 (Owen and Evans, 1998;Owen et al, 2001;Janvier et al, 2003;Doray et al, 2007;Kelly et al, 2008). AP-1A adaptin knock-out's in mice revealed that it is indispensable for mammalian development.…”

Section: Introductionmentioning

confidence: 99%

Characterization of the AP‐1 μ1A and μ1B adaptins in zebrafish (Danio rerio)

Zizioli

Forlanelli

Guarienti

et al. 2010

Developmental Dynamics

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Protein transport between the trans-Golgi network and endosomes is mediated by transport vesicles formed by the adaptor-protein complex AP-1, consisting of the adaptins g1, b1, m1, s1. Mammalia express m1A ubiquitously and isoform m1B in polarized epithelia. Mouse g1 or m1A 'knock out's revealed that AP-1 is indispensable for embryonic development. We isolated m1A and m1B from Danio rerio. Analysis of m1A and m1B expression revealed tissue-specific expression for either one during embryogenesis and in adult tissues in contrast to their expression in mammalia. m1B transcript was detected in organs of endodermal derivation and ''knock-down'' experiments gave rise to embryos defective in formation of intestine, liver, and pronephric ducts. Development ceased at 7-8 dpf. m1B is not expressed in murine liver, indicating loss of m1B expression and establishment of alternative sorting mechanisms during mammalian development. Developmental Dynamics 239:2404-2412,

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A Third Specificity‐Determining Site in μ2 Adaptin for Sequences Upstream of YxxΦ Sorting Motifs

Cited by 47 publications

References 38 publications

Identification of a Non-canonical Tyrosine-based Endocytic Motif in an Ionotropic Receptor

Identification of a Non-canonical Tyrosine-based Endocytic Motif in an Ionotropic Receptor

Structural basis for endosomal trafficking of diverse transmembrane cargos by PX-FERM proteins

Characterization of the AP‐1 μ1A and μ1B adaptins in zebrafish (Danio rerio)

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