Faculty Opinions recommendation of PIP(2)-PDZ domain binding controls the association of syntenin with the plasma membrane.

2002 ◽  
Author(s):  
Craig Montell
Keyword(s):  
Plasma Membrane ◽  
Pdz Domain

scholarly journals The CFTR trafficking mutation F508del inhibits the constitutive activity of SLC26A9

2017 ◽  
Vol 312 (6) ◽  
pp. L912-L925 ◽  
Author(s):  
Carol A. Bertrand ◽  
Shalini Mitra ◽  
Sanjay K. Mishra ◽  
Xiaohui Wang ◽  
Yu Zhao ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Family Member ◽  
Pdz Domain ◽  
Protein S ◽  
Constitutive Activity ◽  
Airway Epithelia ◽  
Anion Secretion ◽  
Anion Transporters ◽  
Bronchial Epithelia ◽  
Domain Protein

Several members of the SLC26A family of anion transporters associate with CFTR, forming complexes in which CFTR and SLC26A functions are reciprocally regulated. These associations are thought to be facilitated by PDZ scaffolding interactions. CFTR has been shown to be positively regulated by NHERF-1, and negatively regulated by CAL in airway epithelia. However, it is unclear which PDZ-domain protein(s) interact with SLC26A9, a SLC26A family member found in airway epithelia. We have previously shown that primary, human bronchial epithelia (HBE) from non-CF donors exhibit constitutive anion secretion attributable to SLC26A9. However, constitutive anion secretion is absent in HBE from CF donors. We examined whether changes in SLC26A9 constitutive activity could be attributed to a loss of CFTR trafficking, and what role PDZ interactions played. HEK293 coexpressing SLC26A9 with the trafficking mutant F508del CFTR exhibited a significant reduction in constitutive current compared with cells coexpressing SLC26A9 and wt CFTR. We found that SLC26A9 exhibits complex glycosylation when coexpressed with F508del CFTR, but its expression at the plasma membrane is decreased. SLC26A9 interacted with both NHERF-1 and CAL, and its interaction with both significantly increased with coexpression of wt CFTR. However, coexpression with F508del CFTR only increased SLC26A9’s interaction with CAL. Mutation of SLC26A9’s PDZ motif decreased this association with CAL, and restored its constitutive activity. Correcting aberrant F508del CFTR trafficking in CF HBE with corrector VX-809 also restored SLC26A9 activity. We conclude that when SLC26A9 is coexpressed with F508del CFTR, its trafficking defect leads to a PDZ motif-sensitive intracellular retention of SLC26A9.

scholarly journals Functional Interaction between Monoamine Plasma Membrane Transporters and the Synaptic PDZ Domain–Containing Protein PICK1

Neuron ◽  
2001 ◽  
Vol 30 (1) ◽  
pp. 121-134 ◽  
Author(s):  
Gonzalo E Torres ◽  
Wei-Dong Yao ◽  
Amy R Mohn ◽  
Hui Quan ◽  
Kyeong-Man Kim ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Pdz Domain ◽  
Membrane Transporters ◽  
Functional Interaction

scholarly journals Formation of a Bazooka–Stardust complex is essential for plasma membrane polarity in epithelia

2010 ◽  
Vol 190 (5) ◽  
pp. 751-760 ◽  
Author(s):  
Michael P. Krahn ◽  
Johanna Bückers ◽  
Lars Kastrup ◽  
Andreas Wodarz
Keyword(s):  
Plasma Membrane ◽  
Protein Complexes ◽  
Pdz Domain ◽  
Phosphorylation Site ◽  
Direct Interaction ◽  
Epithelial Polarity ◽  
Kinase C ◽  
Discs Large ◽  
Evolutionarily Conserved ◽  
Functional Hierarchy

Apical–basal polarity in Drosophila melanogaster epithelia depends on several evolutionarily conserved proteins that have been assigned to two distinct protein complexes: the Bazooka (Baz)–PAR-6 (partitioning defective 6)–atypical protein kinase C (aPKC) complex and the Crumbs (Crb)–Stardust (Sdt) complex. These proteins operate in a functional hierarchy, in which Baz is required for the proper subcellular localization of all other proteins. We investigated how these proteins interact and how this interaction is regulated. We show that Baz recruits Sdt to the plasma membrane by direct interaction between the Postsynaptic density 95/Discs large/Zonula occludens 1 (PDZ) domain of Sdt and a region of Baz that contains a phosphorylation site for aPKC. Phosphorylation of Baz causes the dissociation of the Baz–Sdt complex. Overexpression of a nonphosphorylatable version of Baz blocks the dissociation of Sdt from Baz, causing phenotypes very similar to those of crb and sdt mutations. Our findings provide a molecular mechanism for the phosphorylation-dependent interaction between the Baz–PAR-3 and Crb complexes during the establishment of epithelial polarity.

Several adaptor proteins promote intracellular localisation of the transporter MRP4/ABCC4 in platelets and haematopoietic cells

2017 ◽  
Vol 117 (01) ◽  
pp. 105-115 ◽  
Author(s):  
Yvonne Schaletzki ◽  
Marie-Luise Kromrey ◽  
Susanne Bröderdorf ◽  
Elke Hammer ◽  
Markus Grube ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Protein Interactions ◽  
Pdz Domain ◽  
Adaptor Protein ◽  
Adaptor Proteins ◽  
Apical Plasma Membrane ◽  
Dense Granules ◽  
Haematopoietic Cells ◽  
And Function

SummaryThe multidrug resistance protein 4 (MRP4/ABCC4) has been identified as an important transporter for signalling molecules including cyclic nucleotides and several lipid mediators in platelets and may thus represent a novel target to interfere with platelet function. Besides its localisation in the plasma membrane, MRP4 has been also detected in the membrane of dense granules in resting platelets. In polarised cells it is localised at the basolateral or apical plasma membrane. To date, the mechanism of MRP4 trafficking has not been elucidated; protein interactions may regulate both the localisation and function of this transporter. We approached this issue by searching for interacting proteins by in vitro binding assays, followed by immunoblotting and mass spectrometry, and by visualising their co-localisation in platelets and haematopoietic cells. We identified the PDZ domain containing scaffold proteins ezrin-binding protein 50 (EBP50/NHERF1), postsynaptic density protein 95 (PSD95), and sorting nexin 27 (SNX27), but also the adaptor protein complex 3 subunit β3A (AP3B1) and the heat shock protein HSP90 as putative interaction partners of MRP4. The knockdown of SNX27, PSD95, and AP3B1 by siRNA in megakaryoblastic leuk aemia cells led to a redistribution of MRP4 from intracellular structures to the plasma membrane. Inhibition of HSP90 led to a diminished expression and retention of MRP4 in the endoplasmic reticulum. These results indicate that MRP4 localisation and function are regulated by multiple protein interactions. Changes in the adaptor proteins can hence lead to altered localisation and function of the transporter.Supplementary Material to this article is available at www.thrombosis-online.com.

scholarly journals Syntenin-1 Is a New Component of Tetraspanin-Enriched Microdomains: Mechanisms and Consequences of the Interaction of Syntenin-1 with CD63

2006 ◽  
Vol 26 (20) ◽  
pp. 7707-7718 ◽  
Author(s):  
Nadya Latysheva ◽  
Gairat Muratov ◽  
Sundaresan Rajesh ◽  
Matthew Padgett ◽  
Neil A. Hotchin ◽  
...  
Keyword(s):  
Amino Acids ◽  
Plasma Membrane ◽  
Tyrosine Kinases ◽  
Pdz Domain ◽  
Binding Pocket ◽  
Resonance Spectroscopy ◽  
Nmr Studies ◽  
Transmembrane Receptors ◽  
C Terminus ◽  
Elevated Expression

ABSTRACT Tetraspanins are clustered in specific microdomains (named tetraspanin-enriched microdomains, or TERM) in the plasma membrane and regulate the functions of associated transmembrane receptors, including integrins and receptor tyrosine kinases. We have identified syntenin-1, a PDZ domain-containing protein, as a new component of TERM and show that syntenin-1 specifically interacts with the tetraspanin CD63. Detailed biochemical and heteronuclear magnetic resonance spectroscopy (NMR) studies have demonstrated that the interaction is mediated by the C-terminal cytoplasmic region of the tetraspanin and the PDZ domains of syntenin-1. Upon interaction, NMR chemical shift perturbations were predominantly localized to residues around the binding pocket of PDZ1, indicating a specific mode of recognition of the cytoplasmic tail of CD63. In addition, the C terminus of syntenin-1 has a stabilizing role in the CD63-syntenin-1 association, as deletion of the last 17 amino acids abolished the interaction. The CD63-syntenin-1 complex is abundant on the plasma membrane, and the elevated expression of the wild-type syntenin-1 slows down constitutive internalization of the tetraspanin. Furthermore, internalization of CD63 was completely blocked in cells expressing a syntenin-1 mutant lacking the first 100 amino acids. Previous results have shown that CD63 is internalized via AP-2-dependent mechanisms. Hence, our data indicate that syntenin-1 can counteract the AP-2-dependent internalization and identify this tandem PDZ protein as a new regulator of endocytosis.

Ca2+-dependent inhibition of NHE3 requires PKCα which binds to E3KARP to decrease surface NHE3 containing plasma membrane complexes

2003 ◽  
Vol 285 (6) ◽  
pp. C1527-C1536 ◽  
Author(s):  
Whaseon Lee-Kwon ◽  
Jae Ho Kim ◽  
Jung Woong Choi ◽  
Kazuya Kawano ◽  
Boyoung Cha ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Regulatory Protein ◽  
Pdz Domain ◽  
Dependent Manner ◽  
Null Cell ◽  
Pkc Inhibitor ◽  
Membrane Complex ◽  
Dependent Inhibition ◽  
Intracellular Ca

The intestinal brush border (BB) Na+/H+ exchanger isoform 3 (NHE3) is acutely inhibited by elevation in the concentration of free intracellular Ca2+ ([Ca2+]i) by the cholinergic agonist carbachol and Ca2+ ionophores in a protein kinase C (PKC)-dependent manner. We previously showed that elevating [Ca2+]i with ionomycin rapidly inhibited NHE3 activity and decreased the amount of NHE3 on the plasma membrane in a manner that depended on the presence of the PDZ domain-containing protein E3KARP (NHE3 kinase A regulatory protein, also called NHERF2). The current studies were performed in PS120 fibroblasts (NHE-null cell line) stably transfected with NHE3 and E3KARP to probe the mechanism of PKC involvement in Ca2+ regulation of NHE3. Pretreatment with the general PKC inhibitor, GF109203X prevented ionomycin inhibition of NHE3 without altering basal NHE3 activity. Similarly, the Ca2+-mediated inhibition of NHE3 activity was blocked after pretreatment with the conventional PKC inhibitor Gö-6976 and a specific PKCα pseudosubstrate-derived inhibitor peptide. [Ca2+]i elevation caused translocation of PKCα from cytosol to membrane. PKCα bound to the PDZ1 domain of GST-E3KARP in vitro in a Ca2+-dependent manner. PKCα and E3KARP coimmunoprecipitated from cell lysates; this occurred to a lesser extent at basal [Ca2+]i and was increased with ionomycin exposure. Biotinylation studies demonstrated that [Ca2+]i elevation induced oligomerization of NHE3 in total lysates and decreased the amount of plasma membrane NHE3. Treatment with PKC inhibitors did not affect the oligomerization of NHE3 but did prevent the decrease in surface amount of NHE3. These results suggest that PKCα is not necessary for the Ca2+-dependent formation of the NHE3 plasma membrane complex, although it is necessary for decreasing the membrane amounts of NHE3, probably by stimulating NHE3 endocytosis.

scholarly journals Association of ARVCF with Zonula Occludens (ZO)-1 and ZO-2: Binding to PDZ-Domain Proteins and Cell-Cell Adhesion Regulate Plasma Membrane and Nuclear Localization of ARVCF

2004 ◽  
Vol 15 (12) ◽  
pp. 5503-5515 ◽  
Author(s):  
P. Jaya Kausalya ◽  
Dominic C.Y. Phua ◽  
Walter Hunziker
Keyword(s):  
Plasma Membrane ◽  
Cell Adhesion ◽  
Nuclear Localization ◽  
Pdz Domain ◽  
Cell Contact ◽  
Binding Motif ◽  
Zonula Occludens ◽  
Pdz Domain Proteins ◽  
E Cadherin ◽  
Cell Cell

ARVCF, an armadillo-repeat protein of the p120ctnfamily, associates with classical cadherins and is present in adherens junctions, but its function is poorly understood. Here, we show that ARVCF interacts via a C-terminal PDZ-binding motif with zonula occludens (ZO)-1 and ZO-2. ARVCF and ZO-1 partially colocalize in the vicinity of the apical adhesion complex in polarized epithelial Madin-Darby canine kidney cells. ARVCF, ZO-1, and E-cadherin form a complex and are recruited to sites of initial cell-cell contact in sparse cell cultures. E-cadherin binding and plasma membrane localization of ARVCF require the PDZ-binding motif. Disruption of cell-cell adhesion releases ARVCF from the plasma membrane and an increased fraction of the protein localizes to the nucleus. Nuclear localization of ARVCF also requires the PDZ-binding motif and can be mediated by the PDZ domains of ZO-2. Thus, the interaction of ARVCF with distinct PDZ-domain proteins determines its subcellular localization. Interactions with ZO-1 and ZO-2, in particular, may mediate recruitment of ARVCF to the plasma membrane and the nucleus, respectively, possibly in response to cell-cell adhesion cues.

scholarly journals Drosophila melanogaster Cad99C, the orthologue of human Usher cadherin PCDH15, regulates the length of microvilli

2005 ◽  
Vol 171 (3) ◽  
pp. 549-558 ◽  
Author(s):  
Cecilia D'Alterio ◽  
Dao D.D. Tran ◽  
Maggie W.Y. Au Yeung ◽  
Michael S.H. Hwang ◽  
Michelle A. Li ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Plasma Membrane ◽  
Epithelial Cells ◽  
Binding Sites ◽  
Actin Filaments ◽  
Transmembrane Protein ◽  
Ovarian Follicle ◽  
Pdz Domain ◽  
Follicle Cells ◽  
Apical Surface

Actin-based protrusions can form prominent structures on the apical surface of epithelial cells, such as microvilli. Several cytoplasmic factors have been identified that control the dynamics of actin filaments in microvilli. However, it remains unclear whether the plasma membrane participates actively in microvillus formation. In this paper, we analyze the function of Drosophila melanogaster cadherin Cad99C in the microvilli of ovarian follicle cells. Cad99C contributes to eggshell formation and female fertility and is expressed in follicle cells, which produce the eggshells. Cad99C specifically localizes to apical microvilli. Loss of Cad99C function results in shortened and disorganized microvilli, whereas overexpression of Cad99C leads to a dramatic increase of microvillus length. Cad99C that lacks most of the cytoplasmic domain, including potential PDZ domain–binding sites, still promotes excessive microvillus outgrowth, suggesting that the amount of the extracellular domain determines microvillus length. This study reveals Cad99C as a critical regulator of microvillus length, the first example of a transmembrane protein that is involved in this process.

scholarly journals Confinement of β1- and β2-adrenergic receptors in the plasma membrane of cardiomyocyte-like H9c2 cells is mediated by selective interactions with PDZ domain and A-kinase anchoring proteins but not caveolae

2011 ◽  
Vol 22 (16) ◽  
pp. 2970-2982 ◽  
Author(s):  
Cathleen D. Valentine ◽  
Peter M. Haggie
Keyword(s):  
Plasma Membrane ◽  
Adrenergic Receptors ◽  
Protein Complexes ◽  
Pdz Domain ◽  
Single Particle Tracking ◽  
Live Cells ◽  
Membrane Domains ◽  
Anchoring Proteins ◽  
Differential Signaling ◽  
Receptor Dynamics

The sympathetic nervous system regulates cardiac output by activating adrenergic receptors (ARs) in cardiac myocytes. The predominant cardiac ARs, β1- and β2AR, are structurally similar but mediate distinct signaling responses. Scaffold protein–mediated compartmentalization of ARs into discrete, multiprotein complexes has been proposed to dictate differential signaling responses. To test the hypothesis that βARs integrate into complexes in live cells, we measured receptor diffusion and interactions by single-particle tracking. Unstimulated β1- and β2AR were highly confined in the membrane of H9c2 cardiomyocyte-like cells, indicating that receptors are tethered and presumably integrated into protein complexes. Selective disruption of interactions with postsynaptic density protein 95/disks large/zonula occludens-1 (PDZ)–domain proteins and A-kinase anchoring proteins (AKAPs) increased receptor diffusion, indicating that these scaffold proteins participate in receptor confinement. In contrast, modulation of interactions between the putative scaffold caveolae and β2AR did not alter receptor dynamics, suggesting that these membrane domains are not involved in β2AR confinement. For both β1- and β2AR, the receptor carboxy-terminus was uniquely responsible for scaffold interactions. Our data formally demonstrate that distinct and stable protein complexes containing β1- or β2AR are formed in the plasma membrane of cardiomyocyte-like cells and that selective PDZ and AKAP interactions are responsible for the integration of receptors into complexes.

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