Physiological properties and differential glycosylation of phosphorylated and nonphosphorylated forms of osteopontin secreted by normal rat kidney cells.

In a previous study we have shown that normal rat kidney (NRK) cells in vitro secrete a 69-kDa osteopontin in both phosphorylated (pp69) and nonphosphorylated (np69) forms. Only pp69 interacts with the cell surface and np69 forms a heat-dissociable complex with plasma fibronectin, suggesting functional modulation of osteopontin by phosphorylation. Using tunicamycin, an inhibitor of N-linked glycosylation, and peptide:N-glycosidase F, which removes N-linked oligosaccharide chains from glycoproteins, we show here that np69, but not pp69, contains N-linked carbohydrates. Our results also demonstrate that tunicamycin treatment does not inhibit the cell surface binding of pp69; however, np69 secreted by the treated cells fails to complex with plasma fibronectin, suggesting importantly, our data show that pp69 forms a heat-stable complex with cell surface fibronectin, suggesting that it is an integral component of the extracellular matrix of NRK cells. Finally, sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of deglycosylated and in vitro translated osteopontin suggests that the acidic nature of osteopontin as well as its post-translational modifications play a role in the anomalous behavior of osteopontin in sodium dodecyl sulfate gels, observed in several laboratories. The data presented here provide evidence for possible functional roles of 69-kDa osteopontin and suggest that its physiological properties are regulated by post-translational modifications.

• Publication year

  1990

• Venue

  Journal of Biological Chemistry

• Publication date

  1990-10-25

• Fields of study

  Biology, Medicine, Chemistry

• Identifiers
  DOI 10.1016/s0021-9258(17)44808-3PMID 2211731
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• Normal rat kidney cells secrete both phosphorylated and nonphosphorylated forms of osteopontin showing different physiological properties.

  1989cited by this paper
• Multiple forms of SppI (secreted phosphoprotein, osteopontin) synthesized by normal and transformed rat bone cell populations: regulation by TGF-beta.

  1989cited by this paper
• The nature and significance of osteopontin.

  1989cited by this paper
• Osteopontin, a transformation-associated cell adhesion phosphoprotein, is induced by 12-O-tetradecanoylphorbol 13-acetate in mouse epidermis.

  1989cited by this paper
• Developmental expression of 2ar (osteopontin) and SPARC (osteonectin) RNA as revealed by in situ hybridization

  1988cited by this paper
• Identification of the major phosphoprotein secreted by many rodent cell lines as 2ar/osteopontin: enhanced expression in H-ras-transformed 3T3 cells.

  1988cited by this paper
• Mechanism of fibroblast attachment to bone extracellular matrix: Role of a 44 kilodalton bone phosphoprotein

  1987cited by this paper
• Purification and partial characterization of small proteoglycans I and II, bone sialoproteins I and II, and osteonectin from the mineral compartment of developing human bone.

  1987cited by this paper
• Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction.

  1987cited by this paper
• Isolation, characterization, and biosynthesis of a phosphorylated glycoprotein from rat bone.

  1987cited by this paper
• Tissue specificity and developmental expression of rat osteopontin.

  1987cited by this paper
• 1,25-Dihydroxyvitamin D3 regulates the biosynthesis of osteopontin, a bone-derived cell attachment protein, in clonal osteoblast-like osteosarcoma cells.

  1987cited by this paper
• Molecular cloning of a tumor promoter‐inducible mRNA found in JB6 mouse epidermal cells: Induction is stable at high, but not at low, cell densities

  1987cited by this paper
• Epidermal and transforming growth factors modulate secretion of a 69 kDa phosphoprotein in normal rat kidney fibroblasts

  1987cited by this paper
• New perspectives in cell adhesion: RGD and integrins.

  1987cited by this paper
• Endo-glycosidase F and peptide N-glycosidase F release the great majority of total cellular N-linked oligosaccharides: use in demonstrating that sulfated N-linked oligosaccharides are frequently found in cultured cells.

  1986cited by this paper
• Fibronectins.

  1986cited by this paper
• Isolation and characterization of two sialoproteins present only in bone calcified matrix.

  1985cited by this paper
• Secreted phosphoprotein markers for neoplastic transformation of human epithelial and fibroblastic cells.

  1985cited by this paper
• Altered processing of a major secreted phosphoprotein correlates with tumorigenicity in Rous sarcoma virus-transformed mammalian cells

  1985cited by this paper
• The phosphorylation of proteins: a major mechanism for biological regulation. Fourteenth Sir Frederick Gowland Hopkins memorial lecture.

  1985cited by this paper
• A method for the quantitative recovery of protein in dilute solution in the presence of detergents and lipids.

  1984cited by this paper
• Effect of tunicamycin, an inhibitor of protein glycosylation, on the biological properties of acetylcholine receptor in cultured muscle cells.

  1983cited by this paper
• Binding of plasma fibronectin to cell layers of human skin fibroblasts

  1983cited by this paper
• The EGF receptor-kinase has multiple phosphorylation sites.

  1982cited by this paper
• Cell-free synthesis and glycosylation of the major human-red-cell sialoglycoprotein, glycophorin A.

  1981cited by this paper
• Deglycosylation of glycoproteins by trifluoromethanesulfonic acid.

  1981cited by this paper
• Dimeric character of fibronectin, a major cell surface-associated glycoprotein.

  1977cited by this paper
• Extensive disulfide bonding at the mammalian cell surface.

  1977cited by this paper
• Tunicamycin inhibition of (3H) glucosamine incorporation into yeast glycoproteins: binding of tunicamycin and interaction with phospholipids.

  1976cited by this paper
• Tunicamycin inhibition of polyisoprenyl N-acetylglucosaminyl pyrophosphate formation in calf-liver microsomes.

  1975cited by this paper
• Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4

  1970cited by this paper

• The regulatory role of matrix proteins in mineralization of bone

  2021cites this paper
• Surface-immobilized plant-derived osteopontin as an effective platform to promote osteoblast adhesion and differentiation.

  2019cites this paper
• The role of osteopontin in kidney diseases

  2018cites this paper
• Understanding the Pathophysiology of Nephrocalcinosis

  2017cites this paper
• Osteopontin and vasculogenic mimicry formation are associated with response to neoadjuvant chemotherapy in advanced breast cancer

  2017cites this paper
• Role of osteopontin in lung cancer evolution and heterogeneity.

  2017cites this paper
• Mineralization-inhibiting effects of transglutaminase-crosslinked polymeric osteopontin.

  2017cites this paper
• The Effect of Osteopontin on Microglia

  2017cites this paper
• Investigation of the functional role of osteopontin and its alternatively spliced variants in thyroid carcinoma

  2016cites this paper
• Osteopontin directly modulates cytokine expression of primary microglia and increases their survival.

  2016cites this paper
• Osteopontin Bridging Innate and Adaptive Immunity in Autoimmune Diseases

  2016cites this paper
• Association of OPN rs11730582 polymorphism with cancer risk: a meta-analysis

  2016cites this paper
• Osteopontin mediates survival, proliferation and migration of neural stem cells through the chemokine receptor CXCR4

  2015cites this paper
• Mechanisms of osteopontin-mediated effects on chronic hepatic inflammation and cancer

  2015cites this paper
• Osteopontin Promotes Invasion, Migration and Epithelial-Mesenchymal Transition of Human Endometrial Carcinoma Cell HEC-1A Through AKT and ERK1/2 Signaling

  2015cites this paper
• Probing Protein Assembly, Biomineralization and Biomolecular Interactions by Atomic Force Microscopy

  2014cites this paper
• Osteopontin O-glycosylation contributes to its phosphorylation and cell-adhesion properties.

  2014cites this paper
• High Expression of Genes for Calcificati( in Human Atherosclerotic Plaques on-ri egulating Proteins

  2013cites this paper
• The neuroprotective role of osteopontin in in-vitro and in-vivo models of Parkinson's disease

  2012cites this paper
• О МОДУЛЯТОРАХ ОКСАЛАТНОГО НЕФРОЛИТИАЗА. I. СТИМУЛЯТОРЫ КРИСТАЛЛИЗАЦИИ THE CURRENT CONCEPTIONS ABOUT THE MODULATORS OF THE OXALATE NEPHROLYTHIASIS. I. STIMULATORS OF CRYSTALLIZATION

  2012cites this paper
• Osteopontin, an oxidant stress sensitive cytokine, up‐regulates collagen‐I via integrin αVβ3 engagement and PI3K/pAkt/NFκB signaling

  2012cites this paper
• The role of osteopontin in macrophage-mediated inflammation

  2012cites this paper
• The role of osteopontin and osteopontin aptamer (OPN-R3) in fibroblast activity.

  2012cites this paper
• Elucidating the Role if Integrin-extracellular Matrix Protein Interactions in Regulating Osteoclast Activity

  2011cites this paper
• Expression profile of the matricellular protein osteopontin in primary open-angle glaucoma and the normal human eye.

  2011cites this paper
• Secreted Osteopontin Is Highly Polymerized in Human Airways and Fragmented in Asthmatic Airway Secretions

  2011cites this paper
• High glucose levels increase osteopontin production and pathologic calcification in rat dental pulp tissues.

  2010cites this paper
• Renal Cellular Dysfunction/Damage and the Formation of Kidney Stones

  2010cites this paper
• Selective Calcium Oxalate Crystallization Induced by Monomethylitaconate Grafted Polymethylsiloxane

  2010cites this paper
• Thrombin-cleaved osteopontin regulates hemopoietic stem and progenitor cell functions through interactions with (cid:1) 9 (cid:2) 1 and (cid:1) 4 (cid:2) 1 integrins

  2009cites this paper
• Thrombin-cleaved osteopontin regulates hemopoietic stem and progenitor cell functions through interactions with alpha9beta1 and alpha4beta1 integrins.

  2009cites this paper
• Specifically modified osteopontin in rheumatoid arthritis fibroblast-like synoviocytes supports interaction with B cells and enhances production of interleukin-6.

  2009cites this paper
• Osteopontin and the skin: multiple emerging roles in cutaneous biology and pathology

  2009cites this paper
• Genetic basis of renal cellular dysfunction and the formation of kidney stones

  2009cites this paper
• The role of osteopontin in inflammatory processes

  2009cites this paper
• Fibroblast growth factor‐2 regulates expression of osteopontin in periodontal ligament cells

  2008cites this paper
• Identification of osteopontin phosphorylation sites involved in bone remodeling and inhibition of pathological calcification

  2008cites this paper
• Osteopontin and skeletal muscle myoblasts: association with muscle regeneration and regulation of myoblast function in vitro.

  2008cites this paper
• Post-translational modification and proteolytic processing of urinary osteopontin.

  2008cites this paper
• OSTEOPONTIN AND CELL ADHESION: ROLE OF POST-TRANSLATIONAL MODIFICATIONS AND THE C-TERMINAL REGION by

  2007cites this paper
• Osteopontin: a novel inflammatory mediator of cardiovascular disease.

  2007cites this paper
• SPP1 polymorphisms associated with HBV clearance and HCC occurrence.

  2007cites this paper
• Phosphoproteins of the chicken eggshell calcified layer

  2007cites this paper
• Assembly of collagen-binding peptide with collagen as a bioactive scaffold for osteogenesis in vitro and in vivo.

  2007cites this paper
• Molecular modulation of calcium oxalate crystallization.

  2006cites this paper
• Protein regulation of intrarenal crystallization

  2006cites this paper
• Osteopontin is a downstream effector of the PI3-kinase pathway in melanomas that is inversely correlated with functional PTEN.

  2006cites this paper
• Autocrine secretion of osteopontin results in degradation of IκB in Bcr‐Abl‐expressing cells

  2005cites this paper
• Mycophenolate mofetil prevents the development of experimental autoimmune myocarditis.

  2005cites this paper
• Borrelia burgdorferi, an extracellular pathogen, circumvents osteopontin in inducing an inflammatory cytokine response

  2005cites this paper
• Post-translationally modified residues of native human osteopontin are located in clusters: identification of 36 phosphorylation and five O-glycosylation sites and their biological implications.

  2005cites this paper
• Comprehensive identification of post-translational modifications of rat bone osteopontin by mass spectrometry.

  2005cites this paper
• Influence of macroporous protein scaffolds on bone tissue engineering from bone marrow stem cells.

  2005cites this paper
• Autocrine secretion of osteopontin results in degradation of I kappa B in Bcr-Abl-expressing cells.

  2005cites this paper
• Osteopontin colocalizes with CD44 in cell processes in migrating cells and influences the ligation of CD44 by hyaluronan

  2004cites this paper
• Effects and regulation of osteopontin in rat hepatic stellate cells.

  2004cites this paper
• Expression and characterization of recombinant osteopontin peptides representing matrix metalloproteinase proteolytic fragments.

  2004cites this paper
• Tamm-Horsfall protein is a critical renal defense factor protecting against calcium oxalate crystal formation.

  2004cites this paper
• A locus on chromosome 7 determines dramatic up-regulation of osteopontin in dystrophic cardiac calcification in mice.

  2004cites this paper
• Nuclear Factor-inducing Kinase Plays a Crucial Role in Osteopontin-induced MAPK/IκBα Kinase-dependent Nuclear Factor κB-mediated Promatrix Metalloproteinase-9 Activation*

  2004cites this paper
• Nuclear factor-inducing kinase plays a crucial role in osteopontin-induced MAPK/IkappaBalpha kinase-dependent nuclear factor kappaB-mediated promatrix metalloproteinase-9 activation.

  2004cites this paper
• OSTEOPONTIN EXPRESSION IN PARTICLE-INDUCED LUNG DISEASE

  2004cites this paper
• Identification of a macromolecular crystal growth inhibitor in human urine as osteopontin

  2004cites this paper
• Osteopontin : a protein jthiverse functions

  2004cites this paper
• Osteopontin induces nuclear factor kappa B-mediated promatrix metalloproteinase-2 activation through I kappa B alpha /IKK signaling pathways, and curcumin (diferulolylmethane) down-regulates these pathways.

  2003cites this paper
• The role of osteopontin in lung disease.

  2003cites this paper
• Anti-osteoactivin antibody inhibits osteoblast differentiation and function in vitro.

  2003cites this paper
• Osteopontin Induces Nuclear Factor κB-mediated Promatrix Metalloproteinase-2 Activation through IκBα/IKK Signaling Pathways, and Curcumin (Diferulolylmethane) Down-regulates These Pathways*

  2003cites this paper
• Identification of candidate genes controlling porcine female reproductive traits

  2003cites this paper
• Colocalization of Intracellular Osteopontin With CD44 Is Associated With Migration, Cell Fusion, and Resorption in Osteoclasts

  2002cites this paper
• Mapping of functional epitopes of osteopontin by monoclonal antibodies raised against defined internal sequences

  2002cites this paper
• Characterization of the osteoblast‐like cell phenotype under microgravity conditions in the NASA‐approved rotating wall vessel bioreactor (RWV)

  2002cites this paper
• PDFlib PLOP: PDF Linearization, Optimization, Protection Page inserted by evaluation version

  2001cites this paper
• Renal osteopontin protein and mRNA upregulation during acute nephrotoxicity in the rat.

  2001cites this paper
• Osteopontin Stimulates Tumor Growth and Activation of Promatrix Metalloproteinase-2 through Nuclear Factor-κB-mediated Induction of Membrane Type 1 Matrix Metalloproteinase in Murine Melanoma Cells*

  2001cites this paper
• Expression, roles, receptors, and regulation of osteopontin in the kidney.

  2001cites this paper
• Calcium Oxalate Crystal Attachment to Cultured Rat Kidney Epithelial Cell, NRK-52E

  2001cites this paper
• Apical transport of osteopontin is independent of N-glycosylation and sialylation

  2001influential citation
• Osteopontin: a key cytokine in cell‐mediated and granulomatous inflammation

  2000influential citation
• Phosphorylation of Osteopontin Is Required for Inhibition of Vascular Smooth Muscle Cell Calcification*

  2000cites this paper
• Osteopontin is expressed by adult human osteoarthritic chondrocytes: protein and mRNA analysis of normal and osteoarthritic cartilage.

  2000cites this paper
• Osteopontin expression in fetal and mature human kidney.

  1999cites this paper
• Osteopontin is an autoantigen of the somatostatin cells in human islets: identification by screening random peptide libraries with sera of patients with insulin-dependent diabetes mellitus.

  1999cites this paper
• Proliferation, morphology, and protein expression by osteoblasts cultured on poly(anhydride-co-imides).

  1999cites this paper
• Differences in osteopontin up-regulation between proximal and distal tubules after renal ischemia/reperfusion.

  1999cites this paper
• Alkaline Phosphatase Knock‐Out Mice Recapitulate the Metabolic and Skeletal Defects of Infantile Hypophosphatasia

  1999cites this paper
• [Structural characteristics of osteopontin for calcium oxalate crystal].

  1999cites this paper
• CD44 is not an adhesive receptor for osteopontin

  1999cites this paper
• Regulation of osteopontin gene expression during egg shell formation in the laying hen by mechanical strain.

  1998cites this paper
• Distinct roles of osteopontin in host defense activity and tumor survival during squamous cell carcinoma progression in vivo.

  1998cites this paper
• Differential expression of osteopontin, PC4, and CEC5, a novel mRNA species, during in vitro angiogenesis.

  1998cites this paper
• Urinary Glycoproteins as Modulators of Calcific Nephrolithiasis

  1998cites this paper
• Chapter 14 - Adhesive glycoproteins and receptors

  1997cites this paper
• Osteopontin is the 55-kilodalton fertility-associated protein in Holstein bull seminal plasma.

  1997cites this paper
• Stimulation of osteopontin mRNA expression in HL-60 cells is independent of differentiation.

  1997cites this paper
• Altered sialylation of osteopontin prevents its receptor-mediated binding on the surface of oncogenically transformed tsB77 cells.

  1997cites this paper
• Inhibitors of stone formation.

  1996cites this paper
• The pathogenesis of tubulointerstitial disease associated with glomerulonephritis: the glomerular cytokine theory.

  1995cites this paper
• Gene Expression and Phosphorylation of Mouse Osteopontin a

  1995cites this paper
• Regulation of Osteopontin Expression in Osteoblasts

  1995cites this paper