Home LiteratureArticle Details
PMID: 6358233 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Immunological characterization of the major chick cartilage proteoglycan and its intracellular localization in cultured chondroblasts: a comparison with Type II procollagen.

The Journal of cell biology ·Vol. 97 ·No. 6 ·1983-12-00 ·Pages 1724-36

Pacifici M, Soltesz R, Thal G, Shanley DJ, Boettiger D, Holtzer H

Abstract

Polyclonal antibodies were raised in a rabbit against the major proteoglycan of chick sternal cartilage. A total of six antisera was obtained, three after the first booster injection (A1, A2, and A3) and three after the second booster injection (A4, A5, and A6). The A1 antiserum, which was characterized in most detail, immunoprecipitated native as well as chondroitinase ABC-digested or chondroitinase ABC/keratanase-digested cartilage proteoglycan synthesized by cultured chick chondroblasts, but failed to immunoprecipitate the major proteoglycan synthesized by chick skin fibroblasts. This antiserum was also able to immunoprecipitate the cartilage proteoglycan core protein newly synthesized by cultured chondroblasts, but no other major cell protein. However, the late bleed antisera obtained from the same rabbit after a second booster injection reacted with a new chondroblast-specific polypeptide(s) of approximately 60,000 mol wt in addition to the cartilage proteoglycan. By immunofluorescence procedures, the A1 antiserum stained the extracellular proteoglycan matrix of cultured chondroblasts but not that of skin fibroblasts. Following enzymatic removal of the extracellular matrix and cell membrane permeabilization, this antiserum stained primarily a large, juxtanuclear structure. Additional radioautographic evidence suggests that this structure represents the Golgi complex. Similar immunofluorescent staining with antibodies to the cartilage-characteristic Type II collagen revealed that type II procollagen was localized in numerous cytoplasmic, vacuole-like structures which were scattered throughout most of the chondroblast cytoplasm but were notably scanty in the Golgi complex area. In conclusion, our data suggest the transit of the major cartilage proteoglycan through the Golgi complex of cultured chondroblasts and possible differences in the intracellular distribution of newly synthesized cartilage proteoglycan and Type II procollagen.

MeSH Terms
Animals Antigen-Antibody Complex Cartilage/analysis Cells, Cultured Chickens Fluorescent Antibody Technique Immune Sera Microscopy, Phase-Contrast Procollagen/analysis,immunology Proteoglycans/analysis,immunology
Chemicals
Antigen-Antibody Complex Immune Sera Procollagen Proteoglycans
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Pacifici M
Soltesz R
Thal G
Shanley D J
Boettiger D
Holtzer H
References (65)
65 references, click to expand
  1. Simultaneous localization of type II collagen and core protein of chondroitin sulfate proteoglycan in individual chondrocytes.
    Proc Natl Acad Sci U S A. 1979 Mar;76(3):1261-4 PMID: 375234
  2. Immunohistochemical localization of procollagens. II. Electron microscopic distribution of procollagen I antigenicity in the odontoblasts and predentin of rat incisor teeth by a direct method using peroxidase linked antibodies.
    J Histochem Cytochem. 1979 Jul;27(7):1070-83 PMID: 89154
  3. A comparison of the proteoglycans synthesized in Meckel's and sternal cartilage from normal and nanomelic chick embryos.
    Dev Biol. 1979 Aug;71(2):203-15 PMID: 499656
  4. Translation and characterization of messenger RNAs in differentiating chicken cartilage.
    Proc Natl Acad Sci U S A. 1979 Oct;76(10):4847-51 PMID: 291903
  5. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.
    Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350-4 PMID: 388439
  6. Cell-free synthesis of cartilage proteins: partial identification of proteoglycan core and link proteins.
    Biochemistry. 1980 May 13;19(10):2269-75 PMID: 6246923
  7. 12-O-tetradecanoylphorbol-13-acetate-induced changes in sulfated proteoglycan synthesis in cultured chondroblasts.
    Cancer Res. 1980 Jul;40(7):2461-4 PMID: 7388805
  8. Proteoglycans from chick limb bud chondrocyte cultures. Keratan sulfate and oligosaccharides which contain mannose and sialic acid.
    J Biol Chem. 1980 Jul 10;255(13):6077-83 PMID: 6446553
  9. Oligosaccharides on proteoglycans from the swarm rat chondrosarcoma.
    J Biol Chem. 1980 Jul 10;255(13):6084-91 PMID: 7391006
  10. Structurally distinct collagen types.
    Annu Rev Biochem. 1980;49:957-1003 PMID: 6157354
  11. Physical and immunochemical characterization of proteoglycans synthesized during chondrogenesis in the chick embryo.
    J Biol Chem. 1980 Oct 25;255(20):9870-8 PMID: 7430107
  12. Changes in the sulfated proteoglycans synthesized by "aging" chondrocytes. I. Dispersed cultured chondrocytes and in vivo cartilages.
    J Biol Chem. 1981 Jan 25;256(2):1029-37 PMID: 7451455
  13. Changes in the sulfated proteoglycans synthesized by "aging" chondrocytes. II. Organ-cultured vertebral columns.
    J Biol Chem. 1981 Jan 25;256(2):1038-43 PMID: 7451456
  14. Different proteins associated with 10-nanometer filaments in cultured chick neurons and nonneuronal cells.
    Science. 1981 May 1;212(4494):567-9 PMID: 6163217
  15. Ultrastructure of the chondrocytes and extracellular matrix of the swarm rat chondrosarcoma.
    Anat Rec. 1980 Oct;198(2):135-46 PMID: 7212300
  16. The effect of cycloheximide on synthesis of proteoglycans by cultured chondrocytes from the Swarm rat chondrosarcoma.
    J Biol Chem. 1981 May 10;256(9):4368-76 PMID: 6783661
  17. Identification of core protein, an intermediate in proteoglycan biosynthesis in cultured chondrocytes from the Swarm rat chondrosarcoma.
    J Biol Chem. 1981 Aug 10;256(15):7890-7 PMID: 7263630
  18. Comparison of the NH2-Terminal sequences of chick Type I preprocollagen chains synthesized in an nRNA-dependent reticulocyte lysate.
    Eur J Biochem. 1981 Aug;118(2):363-9 PMID: 6269854
  19. Isolation and characterization of dermatan sulphate and heparan sulphate proteoglycans from fibroblast culture.
    Biochem J. 1981 Jul 1;197(1):217-25 PMID: 6459088
  20. The Golgi apparatus (complex)-(1954-1981)-from artifact to center stage.
    J Cell Biol. 1981 Dec;91(3 Pt 2):77s-103s PMID: 7033246
  21. Purification and structural characterization of a cartilage matrix protein.
    Biochem J. 1981 Aug 1;197(2):367-75 PMID: 7325960
  22. Transformation of chondroblasts by Rous sarcoma virus and synthesis of the sulfated proteoglycan matrix.
    Cell. 1977 Aug;11(4):891-9 PMID: 196766
  23. Chemical and physical changes in proteoglycans during development of chick limb bud chondrocytes grown in vitro.
    J Biol Chem. 1977 Oct 10;252(19):6600-8 PMID: 561071
  24. Persistence of Cartilage proteoglycan and link protein during matrix-induced endochondral bone development: an immunofluorescent study.
    Dev Biol. 1982 Feb;89(2):532-9 PMID: 7035258
  25. Proteochondroitin sulfate synthesis in subcultured chick embryo tibial chondrocytes.
    J Biol Chem. 1982 Feb 25;257(4):1670-5 PMID: 7056736
  26. Isolation and partial characterization of the amino-terminal propeptide of type II procollagen from chick embryos.
    Biochemistry. 1982 Mar 30;21(7):1482-7 PMID: 6177339
  27. Intermediate-size filaments: changes in synthesis and distribution in cells of the myogenic and neurogenic lineages.
    Cold Spring Harb Symp Quant Biol. 1982;46 Pt 1:317-29 PMID: 6955084
  28. A unique low molecular weight collagen secreted by cultured chick embryo chondrocytes.
    J Biol Chem. 1982 Oct 25;257(20):12444-50 PMID: 7118948
  29. Regulation of the synthesis of extracellular matrix components in chondroblasts transformed by a temperature-sensitive mutant of Rous sarcoma virus.
    Cell. 1982 Sep;30(2):373-84 PMID: 6291767
  30. Golgi detection in mitotic and interphase cells by antibodies to secreted galactosyltransferase.
    Exp Cell Res. 1982 Nov;142(1):85-94 PMID: 6814941
  31. Transformation by Rous sarcoma virus induces similar patterns of glycosaminoglycan synthesis in chick embryo skin fibroblasts and vertebral chondroblasts.
    J Biol Chem. 1983 Jan 25;258(2):810-6 PMID: 6296083
  32. Taxol induces microtubule-rough endoplasmic reticulum complexes and microtubule-bundles in cultured chondroblasts.
    Differentiation. 1983;24(1):39-47 PMID: 6135637
  33. Protein measurement with the Folin phenol reagent.
    J Biol Chem. 1951 Nov;193(1):265-75 PMID: 14907713
  34. ON THE SITE OF SULFATION IN THE CHONDROCYTE.
    J Cell Biol. 1964 Jun;21:353-66 PMID: 14189910
  35. STUDIES ON CARTILAGE. V. ELECTRON MICROSCOPIC OBSERVATIONS ON THE AUTORADIOGRAPHIC LOCALIZATION OF S35 IN CELLS AND MATRIX.
    J Ultrastruct Res. 1964 Aug;11:166-72 PMID: 14202313
  36. Electron microscopic and biochemical studies of proteoglycan polydispersity in chick limb bud chondrocyte cultures.
    J Biol Chem. 1978 Jul 10;253(13):4721-9 PMID: 566271
  37. Effects of phorbol-12-myristate-13-acetate on the phenotypic program of cultured chondroblasts and fibroblasts.
    Cancer Res. 1978 Aug;38(8):2350-6 PMID: 667832
  38. Biosynthesis of chondroitin sulfate: microsomal proteoglycans.
    Biochem Biophys Res Commun. 1978 Aug 29;83(4):1502-8 PMID: 697878
  39. Synthesis and extracellular deposition of fibronectin in chondrocyte cultures. Response to the removal of extracellular cartilage matrix.
    J Cell Biol. 1978 Nov;79(2 Pt 1):342-55 PMID: 363726
  40. Characterization of bovine aorta proteoglycan extracted with guanidine hydrochloride in the presence of protease inhibitors.
    J Biol Chem. 1979 Feb 25;254(4):1312-8 PMID: 762130
  41. NH2-terminal sequence of the chick proalpha1(I) chain synthesized in the reticulocyte lysate system. Evidence for a transient hydrophobic leader sequence.
    J Biol Chem. 1979 Mar 10;254(5):1433-6 PMID: 762140
  42. Biosynthesis of proteoglycans by chick limb bud chondrocytes.
    J Biol Chem. 1978 Jul 10;253(13):4713-20 PMID: 566270
  43. Purification and properties of bacterial chondroitinases and chondrosulfatases.
    J Biol Chem. 1968 Apr 10;243(7):1523-35 PMID: 5647268
  44. The loss of phenotypic traits by differentiated cells. VI. Behavior of the progeny of a single chondrocyte.
    J Exp Med. 1969 Aug 1;130(2):417-42 PMID: 5795101
  45. The unlabeled antibody enzyme method of immunohistochemistry: preparation and properties of soluble antigen-antibody complex (horseradish peroxidase-antihorseradish peroxidase) and its use in identification of spirochetes.
    J Histochem Cytochem. 1970 May;18(5):315-33 PMID: 4192899
  46. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  47. Chick cartilage collagen: a new type of alpha 1 chain not present in bone or skin of the species.
    Proc Natl Acad Sci U S A. 1969 Dec;64(4):1264-8 PMID: 5271750
  48. Nascent mucopolysaccharides attached to the Golgi membrane of chondrocytes.
    Biochim Biophys Acta. 1971 Jun 22;237(3):606-10 PMID: 4256144
  49. The specific interaction of hyaluronic acid with cartillage proteoglycans.
    Biochim Biophys Acta. 1972 Sep 15;279(2):401-5 PMID: 4263642
  50. Hyaluronidase digestion and alkaline treatment of bovine tracheal cartilage proteoglycans. Isolation and characterisation of different keratan sulfate proteins.
    Biochim Biophys Acta. 1972 Nov 28;285(1):193-207 PMID: 4265840
  51. Electron microscopic demonstration of proteoglycans in guinea pig epiphyseal cartilage.
    J Ultrastruct Res. 1973 Dec;45(5):407-27 PMID: 4128989
  52. Synthesis, migration, and release of precursor collagen by odontoblasts as visualized by radioautography after (3H)proline administration.
    J Cell Biol. 1974 Jan;60(1):92-127 PMID: 4129079
  53. Ferritin-conjugated antibodies used for labeling of organelles involved in the cellular synthesis and transport of procollagen.
    Proc Natl Acad Sci U S A. 1974 May;71(5):2033-7 PMID: 4525314
  54. Aggregation of cartilage proteoglycans. 3. Characteristics of the proteins isolated from trypsin digests of aggregates.
    J Biol Chem. 1974 Jul 10;249(13):4250-6 PMID: 4277353
  55. Location of procollagen in chick corneal and tendon fibroblasts with ferritin-conjugated antibodies.
    J Cell Biol. 1975 Apr;65(1):75-87 PMID: 1168646
  56. Glycosaminoglycan-synthesizing activity of an isolated Golgi preparation from cultured mast cells.
    Biochem Biophys Res Commun. 1975 Apr 7;63(3):663-8 PMID: 805589
  57. Rapid isolation of antigens from cells with a staphylococcal protein A-antibody adsorbent: parameters of the interaction of antibody-antigen complexes with protein A.
    J Immunol. 1975 Dec;115(6):1617-24 PMID: 1102604
  58. Influence of colchicine and vinblastine on the golgi complex and matrix deposition in chondrocyte aggregates. An ultrastructural study.
    Exp Cell Res. 1975 Oct 15;95(2):440-54 PMID: 1193161
  59. 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):835-51 PMID: 811671
  60. Study of differential collagen synthesis during development of the chick embryo by immunofluorescence. I. Preparation of collagen type I and type II specific antibodies and their application to early stages of the chick embryo.
    Dev Biol. 1976 Feb;48(2):237-49 PMID: 1254083
  61. The route of secretion of procollagen. The influence of alphaalpha'-bipyridyl, colchicine and antimycin A on the secretory process in embryonic-chick tendon and cartilage cells.
    Biochem J. 1976 Apr 15;156(1):81-90 PMID: 8039
  62. Differences among sulfated proteoglycans synthesized in nonchondrogenic cells, presumptive chondroblasts, and chondroblasts.
    Proc Natl Acad Sci U S A. 1976 Sep;73(9):3224-8 PMID: 135259
  63. The occurrence of a new type of proteochondroitin sulfate in the developing chick embryo.
    FEBS Lett. 1976 Dec 1;71(2):337-40 PMID: 1033842
  64. Galactosyl transferase of a Golgi fraction from cultured neoplastic mast cells.
    J Cell Biol. 1977 Mar;72(3):655-66 PMID: 14165
  65. Synthesis of cartilage-specific proteoglycan by suspension cultures of adult chondrocytes.
    Biochem J. 1977 Apr 15;164(1):269-72 PMID: 880233
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.97.6.1724
Published
1983-12-00
Pages
1724-36
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2112721
Subset
IM
Grants
NIA NIH HHS · AG-02758 · United States
NCI NIH HHS · CA-18194 · United States
NHLBI NIH HHS · HL-18708 · United States
External Links
PubMed source DOI: 10.1083/jcb.97.6.1724 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2