Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1989 Oct 1;263(1):105–113. doi: 10.1042/bj2630105

Primary structure of a mouse mastocytoma proteoglycan core protein.

L Kjellén 1, I Pettersson 1, P Lillhager 1, M L Steen 1, U Pettersson 1, P Lehtonen 1, T Karlsson 1, E Ruoslahti 1, L Hellman 1
PMCID: PMC1133396  PMID: 2532501

Abstract

The complete nucleotide sequence of a mouse mastocytoma proteoglycan core protein mRNA was determined. The mRNA, estimated to contain 1.1 kb, encodes a protein with an Mr of 16715. A 21-amino acid-residue region of the protein is composed of alternating serine and glycine residues. Southern-blot analysis of mouse genomic DNA with cDNA containing sequences corresponding to the Ser-Gly repeat region revealed more than 15 gene fragments. Hybridization with a probe corresponding to the N-terminal portion of the core protein identified two fragments, and cDNA covering the C-terminal part of the core protein and the 3' untranslated part of the mRNA hybridized to a single fragment. Antibodies against the core protein, obtained after immunization of rabbits with a fusion protein, reacted with both chondroitin sulphate proteoglycans and heparin proteoglycans produced by the tumour. In immunoblotting of a microsomal fraction from the mastocytoma, the antiserum recognized a single protein (Mr 17,000), which probably represents the core protein before glycosylation.

Full text

PDF
105

Images in this article

108Fig. 2.
on p.108
109Fig. 4.
on p.109
110Fig. 5.
on p.110
110Fig. 6.
on p.110
111Fig. 7.
on p.111

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Avraham S., Stevens R. L., Gartner M. C., Austen K. F., Lalley P. A., Weis J. H. Isolation of a cDNA that encodes the peptide core of the secretory granule proteoglycan of rat basophilic leukemia-1 cells and assessment of its homology to the human analogue. J Biol Chem. 1988 May 25;263(15):7292–7296. [PubMed] [Google Scholar]
  2. BITTER T., MUIR H. M. A modified uronic acid carbazole reaction. Anal Biochem. 1962 Oct;4:330–334. doi: 10.1016/0003-2697(62)90095-7. [DOI] [PubMed] [Google Scholar]
  3. Blobel G., Dobberstein B. Transfer of proteins across membranes. I. Presence of proteolytically processed and unprocessed nascent immunoglobulin light chains on membrane-bound ribosomes of murine myeloma. J Cell Biol. 1975 Dec;67(3):835–851. doi: 10.1083/jcb.67.3.835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bourdon M. A., Krusius T., Campbell S., Schwartz N. B., Ruoslahti E. Identification and synthesis of a recognition signal for the attachment of glycosaminoglycans to proteins. Proc Natl Acad Sci U S A. 1987 May;84(10):3194–3198. doi: 10.1073/pnas.84.10.3194. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bourdon M. A., Oldberg A., Pierschbacher M., Ruoslahti E. Molecular cloning and sequence analysis of a chondroitin sulfate proteoglycan cDNA. Proc Natl Acad Sci U S A. 1985 Mar;82(5):1321–1325. doi: 10.1073/pnas.82.5.1321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bourdon M. A., Shiga M., Ruoslahti E. Gene expression of the chondroitin sulfate proteoglycan core protein PG19. Mol Cell Biol. 1987 Jan;7(1):33–40. doi: 10.1128/mcb.7.1.33. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bourdon M. A., Shiga M., Ruoslahti E. Identification from cDNA of the precursor form of a chondroitin sulfate proteoglycan core protein. J Biol Chem. 1986 Sep 25;261(27):12534–12537. [PubMed] [Google Scholar]
  8. Burnette W. N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. doi: 10.1016/0003-2697(81)90281-5. [DOI] [PubMed] [Google Scholar]
  9. Bywater M., Bywater R., Hellman L. A novel chromatographic procedure for purification of bacterial plasmids. Anal Biochem. 1983 Jul 1;132(1):219–224. doi: 10.1016/0003-2697(83)90451-7. [DOI] [PubMed] [Google Scholar]
  10. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  11. Denhardt D. T. A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Commun. 1966 Jun 13;23(5):641–646. doi: 10.1016/0006-291x(66)90447-5. [DOI] [PubMed] [Google Scholar]
  12. Eccleston E., Leonard B. J., Lowe J. S., Welford H. J. Basophilic leukaemia in the albino rat and a demonstration of the basopoietin. Nat New Biol. 1973 Jul 18;244(133):73–76. doi: 10.1038/newbio244073b0. [DOI] [PubMed] [Google Scholar]
  13. Enerbäck L., Kolset S. O., Kusche M., Hjerpe A., Lindahl U. Glycosaminoglycans in rat mucosal mast cells. Biochem J. 1985 Apr 15;227(2):661–668. doi: 10.1042/bj2270661. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. FURTH J., HAGEN P., HIRSCH E. I. Transplantable mastocytoma in the mouse containing histamine, heparin, 5-hydroxytryptamine. Proc Soc Exp Biol Med. 1957 Aug-Sep;95(4):824–828. doi: 10.3181/00379727-95-23375. [DOI] [PubMed] [Google Scholar]
  15. Hellman L., Pettersson U., Bennich H. Characterization and molecular cloning of the mRNA for the heavy (epsilon) chain of rat immunoglobulin E. Proc Natl Acad Sci U S A. 1982 Feb;79(4):1264–1268. doi: 10.1073/pnas.79.4.1264. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hellman L., Steen M. L., Pettersson U. Nonfunctional immunoglobulin light chain transcripts in two IgE-producing rat immunocytomas; implications for the allelic exclusion and transcription activation processes. Gene. 1985;40(1):115–124. doi: 10.1016/0378-1119(85)90030-7. [DOI] [PubMed] [Google Scholar]
  17. Hök M., Björk I., Hopwood J., Lindahl U. Anticoagulant activity of heparin: separation of high-activity and low-activity heparin species by affinity chromatography on immobilized antithrombin. FEBS Lett. 1976 Jul 1;66(1):90–93. doi: 10.1016/0014-5793(76)80592-3. [DOI] [PubMed] [Google Scholar]
  18. Jacobsson K. G., Riesenfeld J., Lindahl U. Biosynthesis of heparin. Effects of n-butyrate on cultured mast cells. J Biol Chem. 1985 Oct 5;260(22):12154–12159. [PubMed] [Google Scholar]
  19. Katz H. R., Austen K. F., Caterson B., Stevens R. L. Secretory granules of heparin-containing rat serosal mast cells also possess highly sulfated chondroitin sulfate proteoglycans. J Biol Chem. 1986 Oct 15;261(29):13393–13396. [PubMed] [Google Scholar]
  20. Lindahl U., Bäckström G., Thunberg L., Leder I. G. Evidence for a 3-O-sulfated D-glucosamine residue in the antithrombin-binding sequence of heparin. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6551–6555. doi: 10.1073/pnas.77.11.6551. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lindahl U., Pejler G. Heparin-like polysaccharides in intra- and extravascular coagulation reactions. Acta Med Scand Suppl. 1987;715:139–144. doi: 10.1111/j.0954-6820.1987.tb09914.x. [DOI] [PubMed] [Google Scholar]
  22. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  23. Metcalfe D. D., Smith J. A., Austen K. F., Silbert J. E. Polydispersity of rat mast cell heparin. Implications for proteoglycan assembly. J Biol Chem. 1980 Dec 25;255(24):11753–11758. [PubMed] [Google Scholar]
  24. Paulsson M., Sommarin Y., Heinegård D. Metabolism of cartilage proteins in cultured tissue sections. Biochem J. 1983 Jun 15;212(3):659–667. doi: 10.1042/bj2120659. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Robinson H. C., Horner A. A., Hök M., Ogren S., Lindahl U. A proteoglycan form of heparin and its degradation to single-chain molecules. J Biol Chem. 1978 Oct 10;253(19):6687–6693. [PubMed] [Google Scholar]
  26. Ruoslahti E. Structure and biology of proteoglycans. Annu Rev Cell Biol. 1988;4:229–255. doi: 10.1146/annurev.cb.04.110188.001305. [DOI] [PubMed] [Google Scholar]
  27. Schwartz L. B., Austen K. F. Structure and function of the chemical mediators of mast cells. Prog Allergy. 1984;34:271–321. [PubMed] [Google Scholar]
  28. Seldin D. C., Austen K. F., Stevens R. L. Purification and characterization of protease-resistant secretory granule proteoglycans containing chondroitin sulfate di-B and heparin-like glycosaminoglycans from rat basophilic leukemia cells. J Biol Chem. 1985 Sep 15;260(20):11131–11139. [PubMed] [Google Scholar]
  29. Serafin W. E., Katz H. R., Austen K. F., Stevens R. L. Complexes of heparin proteoglycans, chondroitin sulfate E proteoglycans, and [3H]diisopropyl fluorophosphate-binding proteins are exocytosed from activated mouse bone marrow-derived mast cells. J Biol Chem. 1986 Nov 15;261(32):15017–15021. [PubMed] [Google Scholar]
  30. Shively J. E., Conrad H. E. Formation of anhydrosugars in the chemical depolymerization of heparin. Biochemistry. 1976 Sep 7;15(18):3932–3942. doi: 10.1021/bi00663a005. [DOI] [PubMed] [Google Scholar]
  31. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  32. Spindler K. R., Rosser D. S., Berk A. J. Analysis of adenovirus transforming proteins from early regions 1A and 1B with antisera to inducible fusion antigens produced in Escherichia coli. J Virol. 1984 Jan;49(1):132–141. doi: 10.1128/jvi.49.1.132-141.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Stevens R. L., Avraham S., Gartner M. C., Bruns G. A., Austen K. F., Weis J. H. Isolation and characterization of a cDNA that encodes the peptide core of the secretory granule proteoglycan of human promyelocytic leukemia HL-60 cells. J Biol Chem. 1988 May 25;263(15):7287–7291. [PubMed] [Google Scholar]
  34. Stevens R. L., Lee T. D., Seldin D. C., Austen K. F., Befus A. D., Bienenstock J. Intestinal mucosal mast cells from rats infected with Nippostrongylus brasiliensis contain protease-resistant chondroitin sulfate di-B proteoglycans. J Immunol. 1986 Jul 1;137(1):291–295. [PubMed] [Google Scholar]
  35. Stevens R. L., Otsu K., Austen K. F. Purification and analysis of the core protein of the protease-resistant intracellular chondroitin sulfate E proteoglycan from the interleukin 3-dependent mouse mast cell. J Biol Chem. 1985 Nov 15;260(26):14194–14200. [PubMed] [Google Scholar]
  36. Stevens R. L. Secretory granule proteoglycans of mast cells and natural killer cells. Ciba Found Symp. 1986;124:272–285. doi: 10.1002/9780470513385.ch15. [DOI] [PubMed] [Google Scholar]
  37. Tantravahi R. V., Stevens R. L., Austen K. F., Weis J. H. A single gene in mast cells encodes the core peptides of heparin and chondroitin sulfate proteoglycans. Proc Natl Acad Sci U S A. 1986 Dec;83(23):9207–9210. doi: 10.1073/pnas.83.23.9207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Teien A. N., Abildgaard U., Hök M. The anticoagulant effect of heparan sulfate and dermatan sulfate. Thromb Res. 1976 Jun;8(6):859–867. doi: 10.1016/0049-3848(76)90014-1. [DOI] [PubMed] [Google Scholar]
  39. Thunberg L., Bäckström G., Lindahl U. Further characterization of the antithrombin-binding sequence in heparin. Carbohydr Res. 1982 Mar 1;100:393–410. doi: 10.1016/s0008-6215(00)81050-2. [DOI] [PubMed] [Google Scholar]
  40. Woods A., Hök M., Kjellén L., Smith C. G., Rees D. A. Relationship of heparan sulfate proteoglycans to the cytoskeleton and extracellular matrix of cultured fibroblasts. J Cell Biol. 1984 Nov;99(5):1743–1753. doi: 10.1083/jcb.99.5.1743. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. von Heijne G. How signal sequences maintain cleavage specificity. J Mol Biol. 1984 Feb 25;173(2):243–251. doi: 10.1016/0022-2836(84)90192-x. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES