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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Bischoff, F. R. Ponstingl, H. Tilz, G. Maier, G. |
| Description | Author Affiliation: Bischoff FR ( Institute of Cell and Tumor Biology, Deutsches Krebsforschungszentrum, Heidelberg, Federal Republic of Germany.); |
| Abstract | Several autoimmune sera from patients with Raynaud phenomenon decorated mammalian kinetochores and bound to a 47-kDa protein on immunoblots of nuclear lysates. Antibody affinity-purified from immunoblots of the 47-kDa band recognized kinetochores, but due to crossreaction with an 18-kDa protein, localization remains elusive. We used one of these sera to purify the antigen from HeLa cells synchronized in mitosis as a noncovalent complex with a 25-kDa protein. The antigen was released from DNA by intercalation with 25 mM chloroquine. Ion-exchange chromatography yielded the pure complex with an apparent molecular size of 68 kDa, which was separated into its components by gel filtration in 6 M guanidinium chloride. Upon two-dimensional gel electrophoresis the 47-kDa protein gave two main spots of pI 6.6 and 6.7, respectively. Posttranslational modification is indicated by additional antigenic spots, by lack of a free alpha-amino group, and by chromatographic behavior of peptides on reversed-phase chromatography. The amino acid sequence for 205 residues of the 47-kDa protein has been established. This sequence is highly homologous with the translated reading frame of RCC1, a gene reportedly involved in regulating onset of mammalian chromosome condensation. |
| ISSN | 00278424 |
| e-ISSN | 10916490 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Issue Number | 21 |
| Volume Number | 87 |
| Language | English |
| Publisher | National Academy of Sciences |
| Publisher Date | 1990-12-01 |
| Publisher Place | United States |
| Access Restriction | Open |
| Subject Keyword | Autoantibodies Immunology Autoantigens Autoimmune Diseases Chromosomes, Human Physiology Genes Nuclear Proteins Raynaud Disease Amino Acid Sequence Blood Fluorescent Antibody Technique HeLa Cells Molecular Sequence Data Molecular Weight Genetics Sequence Homology, Nucleic Acid Comparative Study Research Support, Non-U.S. Gov't Multidisciplinary |
| Content Type | Text |
| Resource Type | Article |
| Subject | Multidisciplinary |
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