Subnucleosome structures as substrates in enzyme-linked immunosorbent assays
- 5 December 1990
- journal article
- Published by Elsevier BV in Journal of Immunological Methods
- Vol. 134 (2), 187-199
- https://doi.org/10.1016/0022-1759(90)90380-e
Abstract
Histone preparations preserving the tertiary and quaternary structure of histone-histone complexes and histone-DNA complexes, as well as individual histones, were isolated or reconstituted. Various parameters were tested in order to determine the suitability of these complexes for use as substrates in enzyme-linked immunosorbent assays. The protein concentration required to saturate the solid phase was determined, and the amount of bound protein was quantified by the micro-bicinchoninic acid protein assay. In addition, the relative DNA content of solid phase antigen was measured by the binding of monoclonal anti-native DNA antibodies. Prototype sera representing different disease groups produced reproducible and unique patterns of reactivity on the panel of antigens, demonstrating the lack of substantial assay bias. Two substrates, the H2A-H2B dimer and the H2A-H2B-DNA complex, both appear to be oriented in a random manner on the solid phase, leaving a number of different epitopes exposed to the solution. This novel set of histone antigens can now be used to define the specificity of anti-histone antibodies in relation to the quaternary structure of chromatin.Keywords
This publication has 31 references indexed in Scilit:
- Association of Antibody to Histone Complex H2A–H2B with Symptomatic Procainamide-Induced LupusNew England Journal of Medicine, 1988
- Antinuclear antibodies (ANAs): Diagnostically specific immune markers and clues toward the understanding of systemic autoimmunityClinical Immunology and Immunopathology, 1988
- Monoclonal autoantibodies to nuclear antigens from murine graft-versus-host diseaseClinical Immunology and Immunopathology, 1987
- Spectroscopic studies on histone-DNA interactions: II. Three transitions in nucleosomes resolved by salt-titrationJournal of Molecular Biology, 1987
- Spectroscopic studies on histone-DNA interactionsJournal of Molecular Biology, 1987
- Reactivity of anti-histone antibodies induced by procainamide and hydralazineClinical Immunology and Immunopathology, 1982
- Kinetic analysis of deoxyribonuclease I cleavages in the nucleosome core: Evidence for a DNA superhelixJournal of Molecular Biology, 1978
- Purification of the five main calf thymus histone fractions by gel exclusion chromatographyFEBS Letters, 1973
- A new procedure for the isolation and fractionation of histonesFEBS Letters, 1971
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970