scholarly journals Involvement of a cell surface protein and an ecto-protein kinase in myogenesis

1991 ◽  
Vol 279 (2) ◽  
pp. 475-482 ◽  
Author(s):  
X Y Chen ◽  
T C Y Lo
Keyword(s):  
Protein Kinase ◽  
Cell Surface ◽  
Myogenic Differentiation ◽  
Morphological Differentiation ◽  
Surface Protein ◽  
Chemical Reagents ◽  
Defective Mutant ◽  
A Cell ◽  
One Step ◽  
Multinucleated Myotubes

Myogenic differentiation is composed of a sequential cascade of multiple steps leading to the formation of multinucleated myotubes. The interference with any one step would abolish myogenesis. The present investigation examined the cell surface components which might be involved in myogenesis. Studies with subconfluent day 2 cultures of rat L6 myoblasts revealed that a cell surface 112 kDa protein was phosphorylated by a Ca(2+)-, F(-)- and Mg(2+)-dependent ecto-protein kinase [Chen & Lo (1991) Biochem. J. 279, 467-474]. We have shown in the present investigation that adequate ATP was present on the cell surface for efficient functioning of this ecto-protein kinase. The phosphorylation of the 112 kDa protein by this ecto-protein kinase was decrease dramatically in confluent cells and in multinucleated myotubes. The following evidence suggests that both the 112 kDa protein and the ecto-protein kinase may play important roles in myogenesis. (i) The highest phosphorylation activity was observed in subconfluent cultures, i.e. before the onset of morphological differentiation. (ii) Treatment of cells with chemical reagents resulted in a corresponding decrease in the ecto-protein kinase, the 112 kDa protein, the phosphorylated 112 kDa protein (p112) and the ability to form myotubes. (iii) The level of p112 in a conditional myogenesis-defective mutant corresponded with the cells' eventual ability to differentiate. (iv) A mutant defective in the ecto-protein kinase was impaired in the phosphorylation of the 112 kDa protein and in myogenesis. (v) A mutant containing only residual levels of the 112 kDa protein was deficient in both p112 and myogenesis. (vi) Since the level of p112 was normal in another myogenesis-defective mutant, the phosphorylation of this protein was not likely to be a consequence of myogenic differentiation. The above findings suggest that the ecto-protein kinase and the 112 kDa protein may directly or indirectly be associated with the myogenic pathway. Since the levels of the ecto-protein kinase, the 112 kDa protein and p112 decreased dramatically upon the formation of myotubes, these proteins were probably not required once morphological differentiation had been initiated.

Ecto-protein kinase substrate p120 revealed as the cell-surface-expressed nucleolar phosphoprotein Nopp140: a candidate protein for extracellular Ca2+-sensing

2001 ◽  
Vol 360 (3) ◽  
pp. 579-587 ◽  
Author(s):  
Dieter KÜBLER
Keyword(s):  
Protein Kinase ◽  
Cell Surface ◽  
Surface Protein ◽  
Cell Types ◽  
Isolation And Identification ◽  
Kinase Substrate ◽  
Wide Range ◽  
Cell Membrane Proteins ◽  
A Cell ◽  
Mrna Binding

A variety of cell membrane proteins become phosphorylated in their ecto-domains by cell-surface protein kinase (ecto-PK) activities, as detected in a broad spectrum of cell types. This study reports the isolation and identification of a frequent ecto-PK substrate, ecto-p120, using HeLa cells as a model. Data from MS and further biochemical and immunochemical means identified ecto-p120 as a cell-surface homologue of human nucleolar phosphoprotein p140 (hNopp140), which belongs to the family of argyrophilic (AgNOR-stainable) proteins. The superposition of 32P-labelled ecto-nucleolar phosphoprotein p140 (ecto-Nopp140) with anti-Nopp140 immunostaining could be demonstrated in a wide range of cell lines without any exceptions, suggesting a nearly universal occurrence of cell-surface Nopp140. A previous, tentative association of ecto-p120 with the nucleoplasmic pre-mRNA-binding protein hnRNP U has thus been supplanted, since improved purification techniques have allowed unambiguous identification of this ecto-PK cell-surface substrate. Furthermore, we have shown that rapid suppression of ecto-hNopp140 phosphorylation resulted upon a rise in the free extracellular calcium, while lowering the calcium concentrations returned ecto-Nopp140 phosphorylation to the original level. It is important to note that these Ca2+-dependent effects on ecto-Nopp140 phosphorylation are not accompanied by alterations in the phosphorylation of other ecto-PK substrates. Our results indicate that, in addition to nucleolin, a further nucleolar protein, which was considered initially to be strictly intracellular, is identified as a cell-surface phosphoprotein.

scholarly journals Phosphorylation of a cell surface 112 kDa protein by an ecto-protein kinase in rat L6 myoblasts

1991 ◽  
Vol 279 (2) ◽  
pp. 467-474 ◽  
Author(s):  
X Y Chen ◽  
T C Y Lo
Keyword(s):  
Protein Kinase ◽  
Cell Surface ◽  
Myogenic Differentiation ◽  
Protein Profile ◽  
Extracellular Atp ◽  
Experimental Conditions ◽  
Intact Cells ◽  
Low Concentrations ◽  
Km Value ◽  
A Cell

Studies with subconfluent day 2 cultures of rat myoblasts revealed that a cell surface 112 kDa protein could be phosphorylated by extracellular ATP. Analysis of the phosphorylated 112 kDa protein suggested the involvement of a serine protein kinase. The following evidence indicated the cell surface location of this protein kinase: (i) extracellular ATP was unable to penetrate the cell membrane under our experimental conditions; (ii) the phosphorylated protein profile of intact cells differed significantly from that of broken cells; (iii) the phosphorylation of the 112 kDa protein could be abolished by pretreatment of cells with very low concentrations of trypsin; (iv) the phosphorylated 112 kDa protein could be dephosphorylated by exogenously added alkaline phosphatase; (v) the phosphorylation of the 112 kDa protein was inhibited by exogenously added proteins; and (vi) exogenously added proteins could be phosphorylated by intact cells under similar experimental conditions. The phosphorylated 112 kDa protein was detected only when the reaction was carried out in the presence of Ca2+, Mg2+, and F- ions. Kinetic analysis that revealed that the Km value of the ecto-protein kinase for ATP was 0.04 microM, and the Vmax. value for phosphorylation of the 112 kDa protein was 1.67 x 10(-4) pmol/min per 10(5) cells. Data presented in the accompanying paper [Chen & Lo (1991) Biochem. J. 279, 475-482] show that there was a constant and adequate supply of ATP on the cell surface of rat myoblasts for efficient functioning of this protein kinase, and that mutants defective in either the ecto-protein kinase or the 112 kDa protein were also impaired in myogenic differentiation. This and other biochemical studies suggest that the ecto-protein kinase and the 112 kDa protein might play important roles in myogenic differentiation.

scholarly journals Netrins and neogenin promote myotube formation

2004 ◽  
Vol 167 (3) ◽  
pp. 493-504 ◽  
Author(s):  
Jong-Sun Kang ◽  
Min-Jeong Yi ◽  
Wei Zhang ◽  
Jessica L. Feinleib ◽  
Francesca Cole ◽  
...  
Keyword(s):  
Cell Surface ◽  
Myogenic Differentiation ◽  
Surface Proteins ◽  
Myotube Formation ◽  
Skeletal Myoblasts ◽  
Ig Superfamily ◽  
Multistep Process ◽  
A Cell ◽  
Multinucleated Myotubes ◽  
Autocrine Mechanism

Differentiation of skeletal myoblasts into multinucleated myotubes is a multistep process orchestrated by several families of transcription factors, including myogenic bHLH and NFAT proteins. The activities of these factors and formation of myotubes are regulated by signal transduction pathways, but few extracellular factors that might initiate such signals have been identified. One exception is a cell surface complex containing promyogenic Ig superfamily members (CDO and BOC) and cadherins. Netrins and their receptors are established regulators of axon guidance, but little is known of their function outside the nervous system. We report here that myoblasts express the secreted factor netrin-3 and its receptor, neogenin. These proteins stimulate myotube formation and enhance myogenic bHLH- and NFAT-dependent transcription. Furthermore, neogenin binds to CDO in a cis fashion, and myoblasts lacking CDO are defective in responding to recombinant netrin. It is proposed that netrin-3 and neogenin may promote myogenic differentiation by an autocrine mechanism as components of a higher order complex of several promyogenic cell surface proteins.

scholarly journals Leukocytes with chromosome Y loss have reduced abundance of the cell surface immunoprotein CD99

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Jonas Mattisson ◽  
Marcus Danielsson ◽  
Maria Hammond ◽  
Hanna Davies ◽  
Caroline J. Gallant ◽  
...  
Keyword(s):  
Cell Surface ◽  
Single Cells ◽  
Surface Protein ◽  
Protein Abundance ◽  
Blood Leukocytes ◽  
Chromosome Y ◽  
Increased Risk ◽  
A Cell ◽  
Pseudoautosomal Regions ◽  
Male Specific

AbstractMosaic loss of chromosome Y (LOY) in immune cells is a male-specific mutation associated with increased risk for morbidity and mortality. The CD99 gene, positioned in the pseudoautosomal regions of chromosomes X and Y, encodes a cell surface protein essential for several key properties of leukocytes and immune system functions. Here we used CITE-seq for simultaneous quantification of CD99 derived mRNA and cell surface CD99 protein abundance in relation to LOY in single cells. The abundance of CD99 molecules was lower on the surfaces of LOY cells compared with cells without this aneuploidy in all six types of leukocytes studied, while the abundance of CD proteins encoded by genes located on autosomal chromosomes were independent from LOY. These results connect LOY in single cells with immune related cellular properties at the protein level, providing mechanistic insight regarding disease vulnerability in men affected with mosaic chromosome Y loss in blood leukocytes.

Localization and expression of msp130, a primary mesenchyme lineage-specific cell surface protein in the sea urchin embryo

Development ◽  
1987 ◽  
Vol 101 (2) ◽  
pp. 255-265 ◽  
Author(s):  
J.A. Anstrom ◽  
J.E. Chin ◽  
D.S. Leaf ◽  
A.L. Parks ◽  
R.A. Raff
Keyword(s):  
Cell Surface ◽  
Sea Urchin ◽  
Sea Water ◽  
Surface Protein ◽  
Specific Cell ◽  
Cell Surface Protein ◽  
Sea Urchin Embryo ◽  
A Cell ◽  
Mesenchyme Cells ◽  
Primary Mesenchyme Cells

In this report, we use a monoclonal antibody (B2C2) and antibodies against a fusion protein (Leaf et al. 1987) to characterize msp130, a cell surface protein specific to the primary mesenchyme cells of the sea urchin embryo. This protein first appears on the surface of these cells upon ingression into the blastocoel. Immunoelectronmicroscopy shows that msp130 is present in the trans side of the Golgi apparatus and on the extracellular surface of primary mesenchyme cells. Four precursor proteins to msp130 are identified and we show that B2C2 recognizes only the mature form of msp130. We demonstrate that msp130 contains N-linked carbohydrate groups and that the B2C2 epitope is sensitive to endoglycosidase F digestion. Evidence that msp130 is apparently a sulphated glycoprotein is presented. The recognition of the B2C2 epitope of msp130 is disrupted when embryos are cultured in sulphate-free sea water. In addition, two-dimensional immunoblots show that msp130 is an acidic protein that becomes substantially less acidic in the absence of sulphate. We also show that two other independently derived monoclonal antibodies, IG8 (McClay et al. 1983; McClay, Matranga & Wessel, 1985) and 1223 (Carson et al. 1985), recognize msp130, and suggest this protein to be a major cell surface antigen of primary mesenchyme cells.

scholarly journals A cell surface protein controls endocrine ring gland morphogenesis and steroid production

2019 ◽  
Vol 445 (1) ◽  
pp. 16-28 ◽  
Author(s):  
Yanina-Yasmin Pesch ◽  
Ricarda Hesse ◽  
Tariq Ali ◽  
Matthias Behr
Keyword(s):  
Cell Surface ◽  
Surface Protein ◽  
Cell Surface Protein ◽  
Steroid Production ◽  
Ring Gland ◽  
A Cell ◽  
Gland Morphogenesis

scholarly journals The in silico human surfaceome

2018 ◽  
Vol 115 (46) ◽  
pp. E10988-E10997 ◽  
Author(s):  
Damaris Bausch-Fluck ◽  
Ulrich Goldmann ◽  
Sebastian Müller ◽  
Marc van Oostrum ◽  
Maik Müller ◽  
...  
Keyword(s):  
Cell Surface ◽  
In Silico ◽  
Embryonic Stem ◽  
Surface Protein ◽  
Surface Proteins ◽  
Cell Surface Protein ◽  
Cell Surface Proteins ◽  
A Cell ◽  
Visualization Tools ◽  
Protein Classes

Cell-surface proteins are of great biomedical importance, as demonstrated by the fact that 66% of approved human drugs listed in the DrugBank database target a cell-surface protein. Despite this biomedical relevance, there has been no comprehensive assessment of the human surfaceome, and only a fraction of the predicted 5,000 human transmembrane proteins have been shown to be located at the plasma membrane. To enable analysis of the human surfaceome, we developed the surfaceome predictor SURFY, based on machine learning. As a training set, we used experimentally verified high-confidence cell-surface proteins from the Cell Surface Protein Atlas (CSPA) and trained a random forest classifier on 131 features per protein and, specifically, per topological domain. SURFY was used to predict a human surfaceome of 2,886 proteins with an accuracy of 93.5%, which shows excellent overlap with known cell-surface protein classes (i.e., receptors). In deposited mRNA data, we found that between 543 and 1,100 surfaceome genes were expressed in cancer cell lines and maximally 1,700 surfaceome genes were expressed in embryonic stem cells and derivative lines. Thus, the surfaceome diversity depends on cell type and appears to be more dynamic than the nonsurface proteome. To make the predicted surfaceome readily accessible to the research community, we provide visualization tools for intuitive interrogation (wlab.ethz.ch/surfaceome). The in silico surfaceome enables the filtering of data generated by multiomics screens and supports the elucidation of the surfaceome nanoscale organization.

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