Abstract
In this communication is described a new technique for the determination of sedimentation coefficients of macromolecules banded in equilibrium density gradients. Initially, the macromolecules are banded in the analytical ultracentrifuge at a low temperature of about 5°C. After equilibrium has been obtained, the temperature is increased to 25°C. The equilibrium band will now sediment to a new equilibrium position in the ultracentrifuge cell: (a) By following the position of the migrating band as a function of time, sedimentation coefficients may be determined. (b) If several species having different sedimentation coefficients are present in the original band, then during the course of the migration the band may split into several new bands which eventually reunite at the final equilibrium position. (c) If different chemical species of macromolecules such as nucleic acids and carbohydrates are present, in general they will exhibit different temperature density relationships, and can move different distances and directions in response to temperature change.
Full text
PDF
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- COHEN S. S., ARBOGAST R. Chemical studies in host-virus interactions; immunochemical studies on the purity of concentrates of various bacterial viruses prepared by differential centrifugation procedures with an appendix summarizing data on the desoxyribonucleic acid content of these viruses. J Exp Med. 1950 Jun 1;91(6):607–618. doi: 10.1084/jem.91.6.607. [DOI] [PMC free article] [PubMed] [Google Scholar]
- HEARST J. E., IFFT J. B., VINOGRAD J. The effects of pressure on the buoyant behavior of deoxyribonucleic acid and tobacco mosaic virus in a density gradient at equilibrium in the ultracentrifuge. Proc Natl Acad Sci U S A. 1961 Jul 15;47:1015–1025. doi: 10.1073/pnas.47.7.1015. [DOI] [PMC free article] [PubMed] [Google Scholar]
- HEARST J. E. The specific volume of various cationic forms of deoxyribonucleic acid. J Mol Biol. 1962 May;4:415–417. doi: 10.1016/s0022-2836(62)80024-2. [DOI] [PubMed] [Google Scholar]
- HEARST J. E., VINOGRAD J. The net hydration of T-4 bacteriophage deoxyribonuecleic acid and the effect of hydration on buoyant behavior in a density gradient at equilibrium in the ultracentrifuge. Proc Natl Acad Sci U S A. 1961 Jul 15;47:1005–1014. doi: 10.1073/pnas.47.7.1005. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hearst J. E., Vinograd J. A THREE-COMPONENT THEORY OF SEDIMENTATION EQUILIBRIUM IN A DENSITY GRADIENT. Proc Natl Acad Sci U S A. 1961 Jul;47(7):999–1004. doi: 10.1073/pnas.47.7.999. [DOI] [PMC free article] [PubMed] [Google Scholar]
- MANDELL J. D., HERSHEY A. D. A fractionating column for analysis of nucleic acids. Anal Biochem. 1960 Jun;1:66–77. doi: 10.1016/0003-2697(60)90020-8. [DOI] [PubMed] [Google Scholar]
- ROSENBLOOM J., SCHUMAKER V. N. ANALYTICAL ULTRACENTRIFUGATION OF T4R BACTERIOPHAGE DNA IN PREFORMED SUCROSE DENSITY GRADIENTS. Biochemistry. 1963 Nov-Dec;2:1206–1211. doi: 10.1021/bi00906a005. [DOI] [PubMed] [Google Scholar]
- VINOGRAD J., BRUNER R., KENT R., WEIGLE J. Band-centrifugation of macromolecules and viruses in self-generating density gradients. Proc Natl Acad Sci U S A. 1963 Jun;49:902–910. doi: 10.1073/pnas.49.6.902. [DOI] [PMC free article] [PubMed] [Google Scholar]