Generation of truncated forms of the NG2 proteoglycan by cell surface proteolysis. (original) (raw)
- Journal List
- Mol Biol Cell
- v.6(12); 1995 Dec
- PMC301335
Mol Biol Cell. 1995 Dec; 6(12): 1819–1832.
La Jolla Cancer Research Foundation, CA 92037, USA.
Abstract
NG2 is a chondroitin sulfate proteoglycan that is expressed on dividing progenitor cells of several lineages including glia, muscle, and cartilage. It is an integral membrane proteoglycan with a core glycoprotein of 300 kDa. In the present study we have characterized three molecular forms of the NG2 core protein expressed by different cell lines. Many cell lines that express the full length 300-kDa NG2 core protein also release a 290-kDa form into the medium. This species lacks the cytoplasmic domain but contains almost the entire ectodomain. Two core protein species, the intact 300-kDa form and a truncated 275-kDa form, are expressed at the surface of an NG2-transfected cell line U251NG52. The 275-kDa species lacks the cytoplasmic domain and at least 64 amino acids of the ectodomain. Mild trypsinization of B49 cells also generates the 275-kDa species, suggesting that this component is produced by proteolysis of the 300-kDa form. Conversion of the 300-kDa species to the 275-kDa form in U251NG52 cells is stimulated by reagents such as phorbol esters, which activate protein kinase C. Phorbol esters are also known to induce expression of metalloproteinases such as collagenase and stromelysin, which could be responsible for cleavage of the 300-kDa core protein. Although B49 cells do not spontaneously produce the truncated 275-kDa species, use of monoclonal antibodies against NG2 to block the interaction between NG2 and type VI collagen results in the appearance of the 275-kDa component in these cells. Thus the interaction between NG2 and type VI collagen, which contains a Kunitz-type proteinase inhibitor sequence in the alpha 3 chain, may protect the proteoglycan against proteolysis. This is consistent with the observed deficiency of U251NG52 cells in anchoring type VI collagen at the surface.
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- Angel P, Baumann I, Stein B, Delius H, Rahmsdorf HJ, Herrlich P. 12-O-tetradecanoyl-phorbol-13-acetate induction of the human collagenase gene is mediated by an inducible enhancer element located in the 5'-flanking region. Mol Cell Biol. 1987 Jun;7(6):2256–2266. [PMC free article] [PubMed] [Google Scholar]
- Ballin M, Gomez DE, Sinha CC, Thorgeirsson UP. Ras oncogene mediated induction of a 92 kDa metalloproteinase; strong correlation with the malignant phenotype. Biochem Biophys Res Commun. 1988 Aug 15;154(3):832–838. [PubMed] [Google Scholar]
- Bernfield M, Kokenyesi R, Kato M, Hinkes MT, Spring J, Gallo RL, Lose EJ. Biology of the syndecans: a family of transmembrane heparan sulfate proteoglycans. Annu Rev Cell Biol. 1992;8:365–393. [PubMed] [Google Scholar]
- Bialojan C, Takai A. Inhibitory effect of a marine-sponge toxin, okadaic acid, on protein phosphatases. Specificity and kinetics. Biochem J. 1988 Nov 15;256(1):283–290. [PMC free article] [PubMed] [Google Scholar]
- Bonaldo P, Colombatti A. The carboxyl terminus of the chicken alpha 3 chain of collagen VI is a unique mosaic structure with glycoprotein Ib-like, fibronectin type III, and Kunitz modules. J Biol Chem. 1989 Dec 5;264(34):20235–20239. [PubMed] [Google Scholar]
- Bradley RS, Brown AM. The proto-oncogene int-1 encodes a secreted protein associated with the extracellular matrix. EMBO J. 1990 May;9(5):1569–1575. [PMC free article] [PubMed] [Google Scholar]
- Buxbaum JD, Gandy SE, Cicchetti P, Ehrlich ME, Czernik AJ, Fracasso RP, Ramabhadran TV, Unterbeck AJ, Greengard P. Processing of Alzheimer beta/A4 amyloid precursor protein: modulation by agents that regulate protein phosphorylation. Proc Natl Acad Sci U S A. 1990 Aug;87(15):6003–6006. [PMC free article] [PubMed] [Google Scholar]
- Chirgwin JM, Przybyla AE, MacDonald RJ, Rutter WJ. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. [PubMed] [Google Scholar]
- Davis S, Aldrich TH, Ip NY, Stahl N, Scherer S, Farruggella T, DiStefano PS, Curtis R, Panayotatos N, Gascan H, et al. Released form of CNTF receptor alpha component as a soluble mediator of CNTF responses. Science. 1993 Mar 19;259(5102):1736–1739. [PubMed] [Google Scholar]
- DiStefano PS, Chelsea DM, Schick CM, McKelvy JF. Involvement of a metalloprotease in low-affinity nerve growth factor receptor truncation: inhibition of truncation in vitro and in vivo. J Neurosci. 1993 Jun;13(6):2405–2414. [PMC free article] [PubMed] [Google Scholar]
- Downing JR, Roussel MF, Sherr CJ. Ligand and protein kinase C downmodulate the colony-stimulating factor 1 receptor by independent mechanisms. Mol Cell Biol. 1989 Jul;9(7):2890–2896. [PMC free article] [PubMed] [Google Scholar]
- Ehlers MR, Riordan JF. Membrane proteins with soluble counterparts: role of proteolysis in the release of transmembrane proteins. Biochemistry. 1991 Oct 22;30(42):10065–10074. [PubMed] [Google Scholar]
- Flannery CR, Lark MW, Sandy JD. Identification of a stromelysin cleavage site within the interglobular domain of human aggrecan. Evidence for proteolysis at this site in vivo in human articular cartilage. J Biol Chem. 1992 Jan 15;267(2):1008–1014. [PubMed] [Google Scholar]
- Frisch SM, Clark EJ, Werb Z. Coordinate regulation of stromelysin and collagenase genes determined with cDNA probes. Proc Natl Acad Sci U S A. 1987 May;84(9):2600–2604. [PMC free article] [PubMed] [Google Scholar]
- Grako KA, Stallcup WB. Participation of the NG2 proteoglycan in rat aortic smooth muscle cell responses to platelet-derived growth factor. Exp Cell Res. 1995 Nov;221(1):231–240. [PubMed] [Google Scholar]
- Hidaka H, Inagaki M, Kawamoto S, Sasaki Y. Isoquinolinesulfonamides, novel and potent inhibitors of cyclic nucleotide dependent protein kinase and protein kinase C. Biochemistry. 1984 Oct 9;23(21):5036–5041. [PubMed] [Google Scholar]
- Hubbard AL, Cohn ZA. The enzymatic iodination of the red cell membrane. J Cell Biol. 1972 Nov;55(2):390–405. [PMC free article] [PubMed] [Google Scholar]
- Kayyem JF, Roman JM, de la Rosa EJ, Schwarz U, Dreyer WJ. Bravo/Nr-CAM is closely related to the cell adhesion molecules L1 and Ng-CAM and has a similar heterodimer structure. J Cell Biol. 1992 Sep;118(5):1259–1270. [PMC free article] [PubMed] [Google Scholar]
- Lantz M, Gullberg U, Nilsson E, Olsson I. Characterization in vitro of a human tumor necrosis factor-binding protein. A soluble form of a tumor necrosis factor receptor. J Clin Invest. 1990 Nov;86(5):1396–1402. [PMC free article] [PubMed] [Google Scholar]
- Lee MH, Bell RM. The lipid binding, regulatory domain of protein kinase C. A 32-kDa fragment contains the calcium- and phosphatidylserine-dependent phorbol diester binding activity. J Biol Chem. 1986 Nov 15;261(32):14867–14870. [PubMed] [Google Scholar]
- Levine JM. Increased expression of the NG2 chondroitin-sulfate proteoglycan after brain injury. J Neurosci. 1994 Aug;14(8):4716–4730. [PMC free article] [PubMed] [Google Scholar]
- Levine JM, Stallcup WB. Plasticity of developing cerebellar cells in vitro studied with antibodies against the NG2 antigen. J Neurosci. 1987 Sep;7(9):2721–2731. [PMC free article] [PubMed] [Google Scholar]
- Matrisian LM. Metalloproteinases and their inhibitors in matrix remodeling. Trends Genet. 1990 Apr;6(4):121–125. [PubMed] [Google Scholar]
- Mauviel A. Cytokine regulation of metalloproteinase gene expression. J Cell Biochem. 1993 Dec;53(4):288–295. [PubMed] [Google Scholar]
- Mayer U, Pöschl E, Nischt R, Specks U, Pan TC, Chu ML, Timpl R. Recombinant expression and properties of the Kunitz-type protease-inhibitor module from human type VI collagen alpha 3(VI) chain. Eur J Biochem. 1994 Oct 15;225(2):573–580. [PubMed] [Google Scholar]
- Mosley B, Beckmann MP, March CJ, Idzerda RL, Gimpel SD, VandenBos T, Friend D, Alpert A, Anderson D, Jackson J, et al. The murine interleukin-4 receptor: molecular cloning and characterization of secreted and membrane bound forms. Cell. 1989 Oct 20;59(2):335–348. [PubMed] [Google Scholar]
- Nishiyama A, Dahlin KJ, Prince JT, Johnstone SR, Stallcup WB. The primary structure of NG2, a novel membrane-spanning proteoglycan. J Cell Biol. 1991 Jul;114(2):359–371. [PMC free article] [PubMed] [Google Scholar]
- Nishiyama A, Dahlin KJ, Stallcup WB. The expression of NG2 proteoglycan in the developing rat limb. Development. 1991 Apr;111(4):933–944. [PubMed] [Google Scholar]
- Nishiyama A, Stallcup WB. Expression of NG2 proteoglycan causes retention of type VI collagen on the cell surface. Mol Biol Cell. 1993 Nov;4(11):1097–1108. [PMC free article] [PubMed] [Google Scholar]
- Nishizuka Y. The role of protein kinase C in cell surface signal transduction and tumour promotion. Nature. 1984 Apr 19;308(5961):693–698. [PubMed] [Google Scholar]
- Nishizuka Y. The molecular heterogeneity of protein kinase C and its implications for cellular regulation. Nature. 1988 Aug 25;334(6184):661–665. [PubMed] [Google Scholar]
- Pandiella A, Massagué J. Cleavage of the membrane precursor for transforming growth factor alpha is a regulated process. Proc Natl Acad Sci U S A. 1991 Mar 1;88(5):1726–1730. [PMC free article] [PubMed] [Google Scholar]
- Papkoff J, Schryver B. Secreted int-1 protein is associated with the cell surface. Mol Cell Biol. 1990 Jun;10(6):2723–2730. [PMC free article] [PubMed] [Google Scholar]
- Pontén J, Westermark B. Properties of human malignant glioma cells in vitro. Med Biol. 1978 Aug;56(4):184–193. [PubMed] [Google Scholar]
- Prince JT, Milona N, Stallcup WB. Characterization of a partial cDNA clone for the NILE glycoprotein and identification of the encoded polypeptide domain. J Neurosci. 1989 May;9(5):1825–1834. [PMC free article] [PubMed] [Google Scholar]
- Rodriguez-Pena A, Rozengurt E. Disappearance of Ca2+-sensitive, phospholipid-dependent protein kinase activity in phorbol ester-treated 3T3 cells. Biochem Biophys Res Commun. 1984 May 16;120(3):1053–1059. [PubMed] [Google Scholar]
- Sassone-Corsi P, Der CJ, Verma IM. ras-induced neuronal differentiation of PC12 cells: possible involvement of fos and jun. Mol Cell Biol. 1989 Aug;9(8):3174–3183. [PMC free article] [PubMed] [Google Scholar]
- Schrappe M, Klier FG, Spiro RC, Waltz TA, Reisfeld RA, Gladson CL. Correlation of chondroitin sulfate proteoglycan expression on proliferating brain capillary endothelial cells with the malignant phenotype of astroglial cells. Cancer Res. 1991 Sep 15;51(18):4986–4993. [PubMed] [Google Scholar]
- Schubert D, Heinemann S, Carlisle W, Tarikas H, Kimes B, Patrick J, Steinbach JH, Culp W, Brandt BL. Clonal cell lines from the rat central nervous system. Nature. 1974 May 17;249(454):224–227. [PubMed] [Google Scholar]
- Sprecher CA, Kisiel W, Mathewes S, Foster DC. Molecular cloning, expression, and partial characterization of a second human tissue-factor-pathway inhibitor. Proc Natl Acad Sci U S A. 1994 Apr 12;91(8):3353–3357. [PMC free article] [PubMed] [Google Scholar]
- Stallcup WB, Beasley L. Bipotential glial precursor cells of the optic nerve express the NG2 proteoglycan. J Neurosci. 1987 Sep;7(9):2737–2744. [PMC free article] [PubMed] [Google Scholar]
- Stallcup WB, Beasley L, Levine J. Cell-surface molecules that characterize different stages in the development of cerebellar interneurons. Cold Spring Harb Symp Quant Biol. 1983;48(Pt 2):761–774. [PubMed] [Google Scholar]
- Stallcup WB, Dahlin K, Healy P. Interaction of the NG2 chondroitin sulfate proteoglycan with type VI collagen. J Cell Biol. 1990 Dec;111(6 Pt 2):3177–3188. [PMC free article] [PubMed] [Google Scholar]
- Tamaoki T, Nomoto H, Takahashi I, Kato Y, Morimoto M, Tomita F. Staurosporine, a potent inhibitor of phospholipid/Ca++dependent protein kinase. Biochem Biophys Res Commun. 1986 Mar 13;135(2):397–402. [PubMed] [Google Scholar]
- Wilson SS, Baetge EE, Stallcup WB. Antisera specific for cell lines with mixed neuronal and glial properties. Dev Biol. 1981 Apr 15;83(1):146–153. [PubMed] [Google Scholar]
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