Correlated STORM-homoFRET Imaging: Applications to the Study of Membrane Receptor Self-Association and Clustering

Mapping the self-organization and spatial distribution of membrane proteins is key to understanding their function. We report here on a correlated STORM/homoFRET imaging approach for resolving the nanoscale distribution and oligomeric state of membrane proteins. Live cell homoFRET imaging of CEACAM1, a cell-surface receptor known to exist in a complex equilibrium between monomer and dimer/oligomer states, revealed highly heterogenous diffraction-limited structures on the surface of HeLa cells. Correlated super-resolved STORM imaging revealed that these structures comprised a complex mixture and spatial distribution of self-associated CEACAM1 molecules. This correlated approach provides a compelling strategy for addressing challenging questions about the interplay between membrane protein concentration, distribution, interaction, clustering, and function.

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Programmed Cell Death-1 Receptor (PD-1)-Mediated Regulation of Innate Lymphoid Cells

International Journal of Molecular Sciences ◽

10.3390/ijms20112836 ◽

2019 ◽

Vol 20 (11) ◽

pp. 2836 ◽

Cited By ~ 5

Author(s):

Grace Mallett ◽

Arian Laurence ◽

Shoba Amarnath

Keyword(s):

Cell Death ◽

Programmed Cell Death ◽

Innate Lymphoid Cells ◽

Lymphoid Cells ◽

Cell Surface Receptor ◽

Programmed Cell Death 1 ◽

Surface Receptor ◽

A Cell ◽

And Function

Programmed cell death-1 (PD-1) is a cell surface receptor that dampens adaptive immune responses. PD-1 is activated by the engagement of its ligands PDL-1 or PDL-2. This results in the inhibition of T cell proliferation, differentiation, cytokine secretion, and cytolytic function. Although a great deal is known about PD-1 mediated regulation of CD4+ and CD8+ T cells, its expression and function in innate lymphoid cells (ILCs) are yet to be fully deciphered. This review summarizes the role of PD-1 in (1) modulating ILC development, (2) ILC function, and (3) PD-1 signaling in ILC. Finally, we explore how PD-1 based immunotherapies may be beneficial in boosting ILC responses in cancer, infections, and other immune-related disorders.

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RHAMM Is a Centrosomal Protein That Interacts with Dynein and Maintains Spindle Pole Stability

Molecular Biology of the Cell ◽

10.1091/mbc.e02-07-0377 ◽

2003 ◽

Vol 14 (6) ◽

pp. 2262-2276 ◽

Cited By ~ 104

Author(s):

Christopher A. Maxwell ◽

Jonathan J. Keats ◽

Mary Crainie ◽

Xuejun Sun ◽

Tim Yen ◽

...

Keyword(s):

Coiled Coil ◽

Direct Interaction ◽

Spindle Pole ◽

Cell Surface Receptor ◽

Interaction Domain ◽

Surface Receptor ◽

A Cell ◽

Carboxy Terminal ◽

And Function ◽

Hyaluronan Binding

The receptor for hyaluronan-mediated motility (RHAMM), an acidic coiled coil protein, has previously been characterized as a cell surface receptor for hyaluronan, and a microtubule-associated intracellular hyaluronan binding protein. In this study, we demonstrate that a subset of cellular RHAMM localizes to the centrosome and functions in the maintenance of spindle integrity. We confirm a previous study showing that the amino terminus of RHAMM interacts with microtubules and further demonstrate that a separate carboxy-terminal domain is required for centrosomal targeting. This motif overlaps the defined hyaluronan binding domain and bears 72% identity to the dynein interaction domain of Xklp2. RHAMM antibodies coimmunprecipitate dynein IC from Xenopus and HeLa extracts. Deregulation of RHAMM expression inhibits mitotic progression and affects spindle architecture. Structure, localization, and function, along with phylogenetic analysis, suggests that RHAMM may be a new member of the TACC family. Thus, we demonstrate a novel centrosomal localization and mitotic spindle-stabilizing function for RHAMM. Moreover, we provide a potential mechanism for this function in that RHAMM may cross-link centrosomal microtubules, through a direct interaction with microtubules and an association with dynein.

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The 67 kDa laminin receptor: structure, function and role in disease

Bioscience Reports ◽

10.1042/bsr20070004 ◽

2008 ◽

Vol 28 (1) ◽

pp. 33-48 ◽

Cited By ~ 116

Author(s):

John Nelson ◽

Neil V. McFerran ◽

Géraldine Pivato ◽

Emma Chambers ◽

Caroline Doherty ◽

...

Keyword(s):

Sindbis Virus ◽

Eukaryotic Cell ◽

Laminin Receptor ◽

Cell Surface Receptor ◽

Binding Event ◽

Galectin 3 ◽

Surface Receptor ◽

A Cell ◽

Signalling Transduction ◽

And Function

The 67LR (67 kDa laminin receptor) is a cell-surface receptor with high affinity for its primary ligand. Its role as a laminin receptor makes it an important molecule both in cell adhesion to the basement membrane and in signalling transduction following this binding event. The protein also plays critical roles in the metastasis of tumour cells. Isolation of the protein from either normal or cancerous cells results in a product with an approx. molecular mass of 67 kDa. This protein is believed to be derived from a smaller precursor, the 37LRP (37 kDa laminin receptor precursor). However, the precise mechanism by which cytoplasmic 37LRP becomes cell-membrane-embedded 67LR is unclear. The process may involve post-translational fatty acylation of the protein combined with either homo- or hetero-dimerization, possibly with a galectin-3-epitope-containing partner. Furthermore, it has become clear that acting as a receptor for laminin is not the only function of this protein. 67LR also acts as a receptor for viruses, such as Sindbis virus and dengue virus, and is involved with internalization of the prion protein. Interestingly, unmodified 37LRP is a ribosomal component and homologues of this protein are found in all five kingdoms. In addition, it appears to be strongly associated with histones in the eukaryotic cell nucleus, although the precise role of these interactions is not clear. Here we review the current understanding of the structure and function of this molecule, as well as highlighting areas requiring further research.

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Faculty Opinions recommendation of A cell surface receptor mediates extracellular Ca(2+) sensing in guard cells.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1005494.201494 ◽

2004 ◽

Author(s):

Ramón Serrano

Keyword(s):

Cell Surface ◽

Guard Cells ◽

Cell Surface Receptor ◽

Surface Receptor ◽

A Cell

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The transfer of transthyretin and receptor-binding properties from the plasma retinol-binding protein to the epididymal retinoic acid-binding protein

Biochemical Journal ◽

10.1042/bj3620265 ◽

2002 ◽

Vol 362 (2) ◽

pp. 265-271 ◽

Cited By ~ 2

Author(s):

Manickavasagam SUNDARAM ◽

Daan M. F. van AALTEN ◽

John B. C. FINDLAY ◽

Asipu SIVAPRASADARAO

Keyword(s):

Retinoic Acid ◽

Binding Protein ◽

Binding Pocket ◽

Retinol Binding Protein ◽

Cell Surface Receptor ◽

Acid Binding Protein ◽

The Family ◽

Surface Receptor ◽

A Cell ◽

Plasma Retinol

Members of the lipocalin superfamily share a common structural fold, but differ from each other with respect to the molecules with which they interact. They all contain eight β-strands (A—H) that fold to form a well-defined β-barrel, which harbours a binding pocket for hydrophobic ligands. These strands are connected by loops that vary in size and structure and make up the closed and open ends of the pocket. In addition to binding ligands, some members of the family interact with other macromolecules, the specificity of which is thought to be associated with the variable loop regions. Here, we have investigated whether the macromolecular-recognition properties can be transferred from one member of the family to another. For this, we chose the prototypical lipocalin, the plasma retinol-binding protein (RBP) and its close structural homologue the epididymal retinoic acid-binding protein (ERABP). RBP exhibits three molecular-recognition properties: it binds to retinol, to transthyretin (TTR) and to a cell-surface receptor. ERABP binds retinoic acid, but whether it interacts with other macromolecules is not known. Here, we show that ERABP does not bind to TTR and the RBP receptor, but when the loops of RBP near the open end of the pocket (L-1, L-2 and L-3, connecting β-strands A—B, C—D and E—F, respectively) were substituted into the corresponding regions of ERABP, the resulting chimaera acquired the ability to bind TTR and the receptor. L-2 and L-3 were found to be the major determinants of the receptor- and TTR-binding specificities respectively. Thus we demonstrate that lipocalins serve as excellent scaffolds for engineering novel biological functions.

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P4-054: KLOTHO ENHANCES NEURONAL ACTIVITY THROUGH INTERACTION WITH A CELL-SURFACE RECEPTOR

Alzheimer s & Dementia ◽

10.1016/j.jalz.2018.06.2457 ◽

2006 ◽

Vol 14 (7S_Part_27) ◽

pp. P1453-P1454

Author(s):

Nicola J. Corbett ◽

Kate Fisher ◽

Helen A. Rowland ◽

Alys C. Jones ◽

Nigel M. Hooper

Keyword(s):

Cell Surface ◽

Neuronal Activity ◽

Cell Surface Receptor ◽

Surface Receptor ◽

A Cell

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gp96 Is a Human Colonocyte Plasma Membrane Binding Protein for Clostridium difficile Toxin A

Infection and Immunity ◽

10.1128/iai.00326-08 ◽

2008 ◽

Vol 76 (7) ◽

pp. 2862-2871 ◽

Cited By ~ 62

Author(s):

Xi Na ◽

Ho Kim ◽

Mary P. Moyer ◽

Charalabos Pothoulakis ◽

J. Thomas LaMont

Keyword(s):

Plasma Membrane ◽

Clostridium Difficile ◽

Binding Protein ◽

Membrane Binding ◽

Cell Surface Receptor ◽

Toxin A ◽

Surface Receptor ◽

A Cell ◽

Plasma Membrane Binding

ABSTRACT Clostridium difficile toxin A (TxA), a key mediator of antibiotic-associated colitis, requires binding to a cell surface receptor prior to internalization. Our aim was to identify novel plasma membrane TxA binding proteins on human colonocytes. TxA was coupled with biotin and cross-linked to the surface of HT29 human colonic epithelial cells. The main colonocyte binding protein for TxA was identified as glycoprotein 96 (gp96) by coimmunoprecipitation and mass spectrum analysis. gp96 is a member of the heat shock protein family, which is expressed on human colonocyte apical membranes as well as in the cytoplasm. TxA binding to gp96 was confirmed by fluorescence immunostaining and in vitro coimmunoprecipitation. Following TxA binding, the TxA-gp96 complex was translocated from the cell membrane to the cytoplasm. Pretreatment with gp96 antibody decreased TxA binding to colonocytes and inhibited TxA-induced cell rounding. Small interfering RNA directed against gp96 reduced gp96 expression and cytotoxicity in colonocytes. TxA-induced inflammatory signaling via p38 and apoptosis as measured by activation of BAK (Bcl-2 homologous antagonist/killer) and DNA fragmentation were decreased in gp96-deficient B cells. We conclude that human colonocyte gp96 serves as a plasma membrane binding protein that enhances cellular entry of TxA, participates in cellular signaling events in the inflammatory cascade, and facilitates cytotoxicity.

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STRA6, a Cell-Surface Receptor for Retinol-Binding Protein: The Plot Thickens

Cell Metabolism ◽

10.1016/j.cmet.2007.02.006 ◽

2007 ◽

Vol 5 (3) ◽

pp. 164-166 ◽

Cited By ~ 49

Author(s):

William S. Blaner

Keyword(s):

Cell Surface ◽

Binding Protein ◽

Retinol Binding Protein ◽

Cell Surface Receptor ◽

Surface Receptor ◽

A Cell

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230. Megalin, RAP and Nkx3.1 expression in the developing reproductive tract of a marsupial, the tammar wallaby

Reproduction Fertility and Development ◽

10.1071/srb08abs230 ◽

2008 ◽

Vol 20 (9) ◽

pp. 30

Author(s):

M. Gamat ◽

M. B. Renfree ◽

A. J. Pask ◽

G. Shaw

Keyword(s):

Cell Surface ◽

Reproductive Tract ◽

Tammar Wallaby ◽

Cell Surface Receptor ◽

Urogenital Sinus ◽

Receptor Associated Protein ◽

Wide Range ◽

Surface Receptor ◽

A Cell ◽

Strong Staining

Androgens induce the differentiation of the urogenital sinus (UGS) to form a prostate. An early marker of this response is upregulation of the transcription factor Nkx3.1 in the urogenital epithelium in the precursors of prostatic buds. In tammars, prostate differentiation begins ~3 weeks after birth and after the time the testis starts to secrete androgens, and 2 weeks after androgen stimulated Wolffian duct differentiation. The reason for this delay in prostate differentiation is unexplained. Androgen receptors are present in the UGS, and the potent androgen, androstanediol, induces prostatic development in females. Whilst androgens may diffuse into cells by across the cell membrane, there is increasing evidence that steroids are also internalised actively via the cell-surface transport molecule Megalin. We are exploring the possibility that the delay may be related to the establishment of a Megalin-mediated pathway. Megalin is a cell surface receptor expressed on epithelia and mediates the endocytosis of a wide range of ligands, including SHBG-bound sex steroids. Megalin action is regulated by Receptor Associated Protein (RAP), which acts as an antagonist to Megalin action. This study cloned partial sequences of Megalin, RAP and Nkx3.1 and examined their expression in the developing urogenital sinus of the tammar wallaby using RT–PCR. The cellular distribution of Megalin protein in the developing UGS was examined using immunohistochemistry. Megalin, RAP and Nkx3.1 in the tammar were all highly conserved with eutherian orthologueues. Megalin and Nkx3.1 transcripts were detected in the liver, kidney, ovary, testis and developing urogenital sinus of male and female tammars. In the developing UGS of the tammar, there was strong staining for Megalin protein in the urogenital epithelium with some diffuse staining in the surrounding mesenchyme. Together, these results suggest that Megalin could be a key gene in the mediation of androgen action in prostatic development in the tammar wallaby.

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