, Volume 91, Issue 3–4, pp 313–321 | Cite as

Identification of a family of human centromere proteins using autoimmune sera from patients with scleroderma

  • William C. Earnshaw
  • Naomi Rothfield


We have examined “preimmune” serum samples from a patient who progressively developed the symptoms of scleroderma CREST over a period of several years. During this period, anti-centromere antibodies (recognized by indirect immunofluorescence) appeared in the serum. Concomitant with the appearance of the anti-centromere antibodies, antibody species recognizing three chromosomal antigens in immunoblots of SDS polyacrylamide gels appeared in the patient's serum. These antigens migrate with electrophoretic mobilities corresponding to Mr=17, 80, and 140 kilodaltons (kd). Affinity-eluted antibody fractions recognizing the antigens have been prepared from sera of three other patients. Indirect immunofluorescence labeling of mitotic cells using these antibody fractions demonstrates that the antigens are centromere components. We designate them CENP (CENtromere Protein) — A (17kd), CENP-B (80kd), and CENP-C (140kd). The three CENP antigens share antigenic determinants. Immunoblotting experiments show that these patients make antibody species recognizing at least three distinct epitopes on CENP-B and two on CENP-C. Sera from different patients contain different mixtures of the antibody species.


Crest Electrophoretic Mobility Scleroderma Indirect Immunofluorescence Mitotic Cell 
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  1. Bloom KS, Carbon J (1982) Yeast centromere DNA is in a unique and highly ordered structure in chromosomes and small circular minichromosomes. Cell 29:305–317Google Scholar
  2. Burke B, Griffiths G, Louvard D, Roggio H, Warren G (1982) A monoclonal antibody against a 135-K golgi membrane protein. EMBO J 1:1621–1628Google Scholar
  3. Clarke L, Carbon J (1980) Isolation of a yeast centromere and construction of functional small circular minichromosomes. Nature 287:504–509Google Scholar
  4. Clarke L, Carbon J (1983) Genomic substitutions of centromeres in Saccharomyces cerevisiae. Nature 305:23–28Google Scholar
  5. Cox JV, Schenk EA, Olmsted JB (1983) Human anticentromere antibodies: distribution, characterization of antigens, and effect on microtubule organization. Cell 35:331–339Google Scholar
  6. Davis FM, Tsao TY, Fowler SK, Rao PN (1983) Monoclonal antibodies to mitotic cells. Proc Natl Acad Sci USA 80:2926–2930Google Scholar
  7. Douvas AS, Achten M, Tan E (1979) Identification of a nuclear protein (Sc1-70) as a unique target of human antinuclear antibodies in scleroderma. J Biol Chem 254:10514–10522Google Scholar
  8. Earnshaw WC, Laemmli UK (1983) Architecture of metaphase chromosomes and chromosome scaffolds. J Cell Biol 96:84–93Google Scholar
  9. Earnshaw WC, Halligan N, Cooke C, Rothfield N (1984) The kinetochore is part of the metaphase chromosome scaffold. J Cell Biol 98:352–357Google Scholar
  10. Fritzler MJ, Kinsella TD (1980) The CREST syndrome: A distinct serologic entity with anticentromere antibodies. Am J Med 69:520–526Google Scholar
  11. Guldner HH, Lakomek H-J, Bautz FA (1984) Human anti-centromere sera recognise a 19.5 kd nonhistone chromosomal protein from HeLa cells. Clin Exp Immunol (in press)Google Scholar
  12. Lewis CD, Laemmli UK (1982) Higher-order metaphase chromosome structure: evidence for metalloprotein interactions. Cell 29:171–181Google Scholar
  13. McDuffie FC, Bunch TW (1977) Immunologic tests in the diagnosis of rheumatic diseases. Bull Rheum Dis 27:900–905Google Scholar
  14. Moroi Y, Peebles C, Fritzler MJ, Steigerwald J, Tan EM (1980) Autoantibody to centromere (kinetochore) in scleroderma sera. Proc Natl Acad Sci USA 77:1627–1631Google Scholar
  15. Notman DD, Kurata N, Tan EM (1975) Profiles of antinuclear antibodies in systemic rheumatic diseases. Ann Int Med 83:464–469Google Scholar
  16. Olmsted JB (1981) Affinity purification of antibodies from diazotized paper blots of heterogeneous protein samples. J Biol Chem 256:11955–11957Google Scholar
  17. Reichlin M, Mattioli M (1974) Antigens and antibodies characteristic of systemic lupus erythematosus. Bull Rheum Dis 24:756–760Google Scholar
  18. Rieder CL (1982) The mammalian kinetochore. In: Bourne GH, Danielli JF (eds) International review of cytology vol. 79. Academic Press, NY, pp 1–58Google Scholar
  19. Steen VD, Ziegler GL, Rodnan GP, Medsger TA, Jr (1984) Clinical and laboratory associations of anticentromere antibody in patients with progressive systemic sclerosis. Arthritis Rheum 27:125–131Google Scholar
  20. Tan EM, Rodnam GP, Garcia I, Moroi Y, Fritzler MJ, Peebles C (1980) Diversity of antinucclear antibodies in progressive systemic sclerosis. Arthritis Rheum 23:617–625Google Scholar
  21. Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354Google Scholar
  22. Tramposch HD, Smith CD, Senecal J-L, Rothfield N (1984) A long-term longitudinal study of anticentromere antibodies. Arthritis Rheum 27:121–124Google Scholar
  23. Tuffanelli DL, McKeon F, Kleinsmith DM, Burnham TK, Kirschner M (1983) Anticentromere and annticentriole antibodies in the scleroderma spectrum. Arch Dermatol 119:560–566Google Scholar
  24. Williamson DH, Fennell DJ (1975) The use of fluorescent DNA-binding agent for detecting and separating yeast mitochondrial DNA. In: Prescott DM (ed) Methods in cell biology Vol 12. Academic Press, NY, pp 335–351Google Scholar

Copyright information

© Springer-Verlag 1985

Authors and Affiliations

  • William C. Earnshaw
    • 1
  • Naomi Rothfield
    • 2
  1. 1.Department of Cell Biology and AnatomyJohns Hopkins University School of MedicineBaltimoreUSA
  2. 2.Division of Rheumatic Diseases, Department of MedicineUniversity of Connecticut Health CenterFarmingtonUSA

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