Cell-Surface Expression Levels Are Important for Fine-Tuning the Performance of Receptor Tyrosine Kinase-Based Signalobodies

2017 ◽  
Vol 12 (12) ◽  
pp. 1700441
Author(s):  
Hideto Nakabayashi ◽  
Masahiro Kawahara ◽  
Teruyuki Nagamune
Keyword(s):  
Cell Surface ◽  
Tyrosine Kinase ◽  
Receptor Tyrosine Kinase ◽  
Surface Expression ◽  
Fine Tuning ◽  
Cell Surface Expression ◽  
Expression Levels

scholarly journals Position 97 of HLA-B, a residue implicated in pathogenesis of ankylosing spondylitis, plays a key role in cell surface free heavy chain expression

2016 ◽  
Vol 76 (3) ◽  
pp. 593-601 ◽  
Author(s):  
Liye Chen ◽  
Hui Shi ◽  
Jack Yuan ◽  
Paul Bowness
Keyword(s):  
Ankylosing Spondylitis ◽  
Cell Surface ◽  
Heavy Chain ◽  
Protein Interactions ◽  
Hela Cells ◽  
Antigen Processing ◽  
Surface Expression ◽  
Cell Surface Expression ◽  
Expression Levels ◽  
Free Heavy Chain

ObjectiveAssociation of position 97 (P97) residue polymorphisms in human leucocyte antigen (HLA)-B, including HLA-B*27, with ankylosing spondylitis (AS) has recently been reported. We studied the effect of P97 variations on cell surface expression of the AS-associated HLA-B*27 and HLA-B*51, and the AS-protective HLA-B*7.MethodsFlow cytometry was used to measure surface expression of HLA-B*27 in C1R/HeLa cells expressing HLA-B*27 (N97) and six mutants at P97 (N97T, N97S, N97V, N97R, N97W and N97D). Transporter associated with antigen processing-deficient T2, tapasin-deficient 220, β2m-deficient HCT15 and endoplasmic reticulum aminopeptidase 1 or β2m-clustered regularly interspaced short palindromic repeats/Cas9-knockout HeLa cells were used to provide evidence for specific protein interactions. Surface expression of HLA-B*7/HLA-B*51 P97 mutants was also studied.ResultsMutation of HLA-B*27 P97 to the AS risk residue threonine increased cell surface free heavy chain (FHC) expression. Protective residues (serine or valine) and non-AS-associated residues (arginine or tryptophan) did not alter FHC expression. The N97D mutation reduced expression of conventional and FHC forms of HLA-B*27. Differences in FHC expression levels between HLA-B*27, HLA-B*27-N97T and HLA-B*27-N97D were dependent on the presence of functional β2m. HLA-B*7, which has an AS-protective serine at P97, expressed lower levels of FHC than HLA-B*27 or HLA-B*51. Introduction of asparagine at P97 of both HLA-B*7 and HLA-B*51 increased FHC expression.ConclusionsThe nature of P97 residue affects surface expression of HLA-B*27, B*7 and B*51, with AS-associated residues giving rise to higher FHC expression levels. The association of P97 amino acid polymorphisms with AS could be, at least in part, explained by its effect on HLA-B*27 FHC cell surface expression.

scholarly journals Trafficking of Kv1.4 potassium channels: interdependence of a pore region determinant and a cytoplasmic C-terminal VXXSL determinant in regulating cell-surface trafficking

2003 ◽  
Vol 375 (3) ◽  
pp. 761-768 ◽  
Author(s):  
Jing ZHU ◽  
Itaru WATANABE ◽  
Barbara GOMEZ ◽  
William B. THORNHILL
Keyword(s):  
Cell Surface ◽  
Cell Function ◽  
Cell Signalling ◽  
Surface Protein ◽  
Surface Expression ◽  
Inhibitory Effect ◽  
Cell Surface Expression ◽  
Pore Region ◽  
Expression Levels ◽  
Protein Levels

Kv1.4 and Kv1.1 potassium channel homomers have been shown to exhibit different intracellular trafficking programmes and cell-surface expression levels in cell lines: a determinant in the pore region of Kv1.4 and Kv1.1 [Zhu, Watanabe, Gomez and Thornhill (2001) J. Biol. Chem. 276, 39419–39427] and a cytoplasmic C-terminal VXXSL determinant on Kv1.4 [Li, Takimoto and Levitan (2000) J. Biol. Chem. 275, 11597–11602] have been described, which affected trafficking and cell-surface expression levels. In the present study, we examined whether trafficking pore determinants influenced any cytoplasmic C-terminal trafficking determinant. We found that removal of VXXSL from a Kv1.4 chimaera that contained the pore of Kv1.1 did not affect cell-surface trafficking. Therefore removal of the C-terminal VXXSL of Kv1.4 inhibited protein surface levels only in the presence of the Kv1.4 pore. In contrast, truncating the cytoplasmic C-terminus of Kv1.1 or truncating a Kv1.1 chimaera with the pore of Kv1.4, had little effect on surface protein levels. Furthermore, the subregion of the Kv1.4 pore trafficking determinant that was required for the inhibitory effect of VXXSL removal was mapped to a threonine residue in the deep pore region. Therefore the Kv1.4 pore determinant affected the trafficking and cell-surface levels directed by the C-terminal VXXSL determinant. Different Kv1 trafficking programmes would affect cell-surface expression levels either positively or negatively and also cell signalling. Cells may use differential trafficking programmes of membrane proteins as a post-translational mechanism to regulate surface protein levels and cell function.

scholarly journals DNA methylation regulates adenosine A2Areceptor cell surface expression levels

2010 ◽  
Vol 112 (5) ◽  
pp. 1273-1285 ◽  
Author(s):  
Sandra P. Buira ◽  
José Luis Albasanz ◽  
Guido Dentesano ◽  
Jesús Moreno ◽  
Mairena Martín ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Cell Surface ◽  
Surface Expression ◽  
Cell Surface Expression ◽  
Expression Levels

scholarly journals DC-SIGN Interactions with Human Immunodeficiency Virus Type 1 and 2 and Simian Immunodeficiency Virus

2001 ◽  
Vol 75 (10) ◽  
pp. 4664-4672 ◽  
Author(s):  
Stefan Pöhlmann ◽  
Frédéric Baribaud ◽  
Benhur Lee ◽  
George J. Leslie ◽  
Melissa D. Sanchez ◽  
...  
Keyword(s):  
Human Immunodeficiency Virus ◽  
Cell Surface ◽  
Lectin Binding ◽  
Surface Expression ◽  
Cell Surface Expression ◽  
Virus Binding ◽  
Expression Levels ◽  
Immunodeficiency Virus ◽  
Hiv 1

ABSTRACT Dendritic cells (DCs) efficiently bind and transmit human immunodeficiency virus (HIV) to cocultured T cells and so may play an important role in HIV transmission. DC-SIGN, a novel C-type lectin that is expressed in DCs, has recently been shown to bind R5 HIV type 1 (HIV-1) strains and a laboratory-adapted X4 strain. To characterize the interaction of DC-SIGN with primate lentiviruses, we investigated the structural determinants of DC-SIGN required for virus binding and transmission to permissive cells. We constructed a panel of DC-SIGN mutants and established conditions which allowed comparable cell surface expression of all mutants. We found that R5, X4, and R5X4 HIV-1 isolates as well as simian immunodeficiency and HIV-2 strains bound to DC-SIGN and could be transmitted to CD4/coreceptor-positive cell types. DC-SIGN contains a single N-linked carbohydrate chain that is important for efficient cell surface expression but is not required for DC-SIGN-mediated virus binding and transmission. In contrast, C-terminal deletions removing either the lectin binding domain or the repeat region abrogated DC-SIGN function. Trypsin-EDTA treatment inhibited DC-SIGN mediated infection, indicating that virus was maintained at the surface of the DC-SIGN-expressing cells used in this study. Finally, quantitative fluorescence-activated cell sorting analysis of AU1-tagged DC-SIGN revealed that the efficiency of virus transmission was strongly affected by variations in DC-SIGN expression levels. Thus, variations in DC-SIGN expression levels on DCs could greatly affect the susceptibility of human individuals to HIV infection.

scholarly journals Regulation of TrkB cell surface expression—a mechanism for modulation of neuronal responsiveness to brain-derived neurotrophic factor

2020 ◽  
Vol 382 (1) ◽  
pp. 5-14 ◽  
Author(s):  
Thomas Andreska ◽  
Patrick Lüningschrör ◽  
Michael Sendtner
Keyword(s):  
Cell Surface ◽  
Neurotrophic Factor ◽  
Tyrosine Kinases ◽  
Receptor Tyrosine Kinases ◽  
Surface Expression ◽  
Brain Derived Neurotrophic Factor ◽  
Fine Tuning ◽  
Cell Surface Expression ◽  
Rapid Modulation ◽  
And Function

Abstract Neurotrophin signaling via receptor tyrosine kinases is essential for the development and function of the nervous system in vertebrates. TrkB activation and signaling show substantial differences to other receptor tyrosine kinases of the Trk family that mediate the responses to nerve growth factor and neurotrophin-3. Growing evidence suggests that TrkB cell surface expression is highly regulated and determines the sensitivity of neurons to brain-derived neurotrophic factor (BDNF). This translocation of TrkB depends on co-factors and modulators of cAMP levels, N-glycosylation, and receptor transactivation. This process can occur in very short time periods and the resulting rapid modulation of target cell sensitivity to BDNF could represent a mechanism for fine-tuning of synaptic plasticity and communication in complex neuronal networks. This review focuses on those modulatory mechanisms in neurons that regulate responsiveness to BDNF via control of TrkB surface expression.

scholarly journals Allelic variation of killer cell immunoglobulin-like receptor 2DS5 impacts glycosylation altering cell surface expression levels

2014 ◽  
Vol 75 (2) ◽  
pp. 124-128 ◽  
Author(s):  
Noriko K. Steiner ◽  
Sivanesan Dakshanamurthy ◽  
Nicholas Nguyen ◽  
Carolyn Katovich Hurley
Keyword(s):  
Cell Surface ◽  
Allelic Variation ◽  
Killer Cell ◽  
Surface Expression ◽  
Cell Surface Expression ◽  
Expression Levels

The Degree of N-glycosylation Affects the Trafficking and Cell Surface Expression Levels of Kv1.4 Potassium Channels

2014 ◽  
Vol 248 (2) ◽  
pp. 187-196 ◽  
Author(s):  
Itaru Watanabe ◽  
Jing Zhu ◽  
Esperanza Recio-Pinto ◽  
William B. Thornhill
Keyword(s):  
Potassium Channels ◽  
Cell Surface ◽  
Surface Expression ◽  
Cell Surface Expression ◽  
Expression Levels
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