Abstract
The spectrum of cellular targets in the autoimmune diseases is both large and varied and includes among the nuclear components the so-called Sm and RNP antigens associated with systemic lupus erythematosus. The use of immunoaffinity chromatography with dual specificity for the Sm and RNP antigens has allowed for their substantial purification from rabbit thymus in parallel and in quantity. In lieu of a functional assay, the use of a counterimmunoelectrophoresis assay provided a sensitive and rapid means of monitoring the distribution of the two antigens during purification and ensured the isolation of complexes containing the components required for antigenicity. The resulting purified complex consisted of nine polypeptides having molecular weights of approximately 9000 to 44,000 and two small RNAs of similar size. However, limited proteolysis of the isolated complex suggested that most of these polypeptides were not actually required for antigenic activity. Unlike Sm in crude thymus extracts, purified Sm was RNase sensitive. Thus, one of the major diagnostic criteria used to distinguish Sm and RNP antigens in crude extracts was shown to be invalid for purified material, suggesting that both antigens from rabbit thymus are actually ribonucleoprotein complexes.
Full text
PDFImages in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Blobel G., Potter V. R. Nuclei from rat liver: isolation method that combines purity with high yield. Science. 1966 Dec 30;154(3757):1662–1665. doi: 10.1126/science.154.3757.1662. [DOI] [PubMed] [Google Scholar]
- Chandler P. M., Rimkus D., Davidson N. Gel electrophoretic fractionation of RNAs by partial denaturation with methylmercuric hydroxide. Anal Biochem. 1979 Oct 15;99(1):200–206. doi: 10.1016/0003-2697(79)90063-0. [DOI] [PubMed] [Google Scholar]
- Dahlberg A. E., Dingman C. W., Peacock A. C. Electrophoretic characterization of bacterial polyribosomes in agarose-acrylamide composite gels. J Mol Biol. 1969 Apr 14;41(1):139–147. doi: 10.1016/0022-2836(69)90131-4. [DOI] [PubMed] [Google Scholar]
- Douvas A. S., Stumph W. E., Reyes P., Tan E. M. Isolation and characterization of nuclear ribonucleoprotein complexes using human anti-nuclear ribonucleoprotein antibodies. J Biol Chem. 1979 May 10;254(9):3608–3616. [PubMed] [Google Scholar]
- Fairbanks G., Steck T. L., Wallach D. F. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 1971 Jun 22;10(13):2606–2617. doi: 10.1021/bi00789a030. [DOI] [PubMed] [Google Scholar]
- Franco-Saenz R., Suzuki S., Tan S. Y. Prostaglandins and renin production: a review. Prostaglandins. 1980 Dec;20(6):1131–1143. doi: 10.1016/0090-6980(80)90065-9. [DOI] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Lerner M. R., Boyle J. A., Mount S. M., Wolin S. L., Steitz J. A. Are snRNPs involved in splicing? Nature. 1980 Jan 10;283(5743):220–224. doi: 10.1038/283220a0. [DOI] [PubMed] [Google Scholar]
- Lerner M. R., Steitz J. A. Antibodies to small nuclear RNAs complexed with proteins are produced by patients with systemic lupus erythematosus. Proc Natl Acad Sci U S A. 1979 Nov;76(11):5495–5499. doi: 10.1073/pnas.76.11.5495. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mattioli M., Reichlin M. Physical association of two nuclear antigens and mutual occurrence of their antibodies: the relationship of the SM and RNAprotein (MO) systems in SLE sera. J Immunol. 1973 May;110(5):1318–1324. [PubMed] [Google Scholar]
- Nakamura R. M., Tan E. M. Recent progress in the study of autoantibodies to nuclear antigens. Hum Pathol. 1978 Jan;9(1):85–91. doi: 10.1016/s0046-8177(78)80010-0. [DOI] [PubMed] [Google Scholar]
- Notman D. D., Kurata N., Tan E. M. Profiles of antinuclear antibodies in systemic rheumatic diseases. Ann Intern Med. 1975 Oct;83(4):464–469. doi: 10.7326/0003-4819-83-4-464. [DOI] [PubMed] [Google Scholar]
- Peltier A. P., Aussel C., Haim T., Cyna L. Ribonucleoprotein (RNP) soluble nuclear antigen: demonstration of its reaction with serum antibodies by counter-immunoelectrophoresis and further partial characterization. J Immunol Methods. 1977;16(2):153–163. doi: 10.1016/0022-1759(77)90050-3. [DOI] [PubMed] [Google Scholar]
- Raj N. B., Ro-Choi T. S., Busch H. Nuclear ribonucleoprotein complexes containing U1 and U2 RNA. Biochemistry. 1975 Oct 7;14(20):4380–4385. doi: 10.1021/bi00691a006. [DOI] [PubMed] [Google Scholar]
- Sharp G. C., Irvin W. S., May C. M., Holman H. R., McDuffie F. C., Hess E. V., Schmid F. R. Association of antibodies to ribonucleoprotein and Sm antigens with mixed connective-tissue disease, systematic lupus erythematosus and other rheumatic diseases. N Engl J Med. 1976 Nov 18;295(21):1149–1154. doi: 10.1056/NEJM197611182952101. [DOI] [PubMed] [Google Scholar]
- Shibata H., Ro-Choi T. S., Reddy R., Choi Y. C., Henning D., Busch H. The primary nucleotide sequence of nuclear U-2 ribonucleic acid. The 5'-terminal portion of the molecule. J Biol Chem. 1975 May 25;250(10):3909–3920. [PubMed] [Google Scholar]
- Waelti E. R., Hess M. W. Nuclear antigens: characterization of the acidic nuclear ribonucleoprotein-Sm-antigen complex by analytical and preparative isoelectric focusing. J Immunol Methods. 1980;32(2):177–184. doi: 10.1016/0022-1759(80)90071-x. [DOI] [PubMed] [Google Scholar]
- Wilchek M., Bocchini V., Becker M., Givol D. A general method for the specific isolation of peptides containing modified residues, using insoluble antibody columns. Biochemistry. 1971 Jul 20;10(15):2828–2834. doi: 10.1021/bi00791a004. [DOI] [PubMed] [Google Scholar]