Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
Bischoff, FR; Maier, G; Tilz, G; Ponstingl, H.
A 47-kDa human nuclear protein recognized by antikinetochore autoimmune sera is homologous with the protein encoded by RCC1, a gene implicated in onset of chromosome condensation.
PROC NAT ACAD SCI USA. 1990; 87(21): 8617-8621.
Doi: 10.1073/pnas.87.21.8617
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- Co-Autor*innen der Med Uni Graz
- Tilz Gernot
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- 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.
- Find related publications in this database (using NLM MeSH Indexing)
- Amino Acid Sequence -
- Autoantibodies - immunology
- Autoantigens - analysis
- Autoimmune Diseases - blood
- Chromosomes, Human - physiology
- Female - physiology
- Fluorescent Antibody Technique - physiology
- Genes - physiology
- Hela Cells - immunology
- Humans - immunology
- Middle Aged - immunology
- Molecular Sequence Data - immunology
- Molecular Weight - immunology
- Nuclear Proteins - analysis
- Raynaud Disease - blood
- Sequence Homology, Nucleic Acid - blood