Abstract
Sedimentation equilibrium measurements can be used to determine the molecular weight of the protein moiety of a protein-detergent complex without prior knowledge of detergent binding. The procedure is to adjust the solvent density by addition of D2O so as to blank out the contribution of bound detergent to the sedimentation potential. An approximate measure of detergent binding can be obtained from the effect of solvent density on the sedimentation result. The procedure is also applicable to protein-lipid complexes. It can be used for complexes containing both lipid and detergent if the lipid content is known. The use of the method is demonstrated by experimental data for the AI polypeptide of serum high density lipoprotein, in separate complexes with nonionic detergents and with a phospholipid.
Full text
PDF



Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Baker H. N., Gotto A. M., Jr, Jackson R. L. The primary structure of human plasma high density apolipoprotein glutamine I (ApoA-I). II. The amino acid sequence and alignment of cyanogen bromide fragments IV, III, and I. J Biol Chem. 1975 Apr 10;250(7):2725–2738. [PubMed] [Google Scholar]
- CASASSA E. F., EISENBERG H. THERMODYNAMIC ANALYSIS OF MULTICOMPONENT SOLUTIONS. Adv Protein Chem. 1964;19:287–395. doi: 10.1016/s0065-3233(08)60191-6. [DOI] [PubMed] [Google Scholar]
- Edelstein S. J., Schachman H. K. The simultaneous determination of partial specific volumes and molecular weights with microgram quantities. J Biol Chem. 1967 Jan 25;242(2):306–311. [PubMed] [Google Scholar]
- Fisher W. R., Granade M. E., Mauldin J. L. Hydrodynamic studies of human low density lipoproteins. Evaluation of the diffusion coefficient and the preferential hydration. Biochemistry. 1971 Apr 27;10(9):1622–1629. doi: 10.1021/bi00785a019. [DOI] [PubMed] [Google Scholar]
- Friedberg S. J., Reynolds J. A. The molar ratio of the two major polypeptide components of human high density lipoprotein. J Biol Chem. 1976 Jul 10;251(13):4005–4009. [PubMed] [Google Scholar]
- Grefrath S. P., Reynolds J. A. The molecular weight of the major glycoprotein from the human erythrocyte membrane. Proc Natl Acad Sci U S A. 1974 Oct;71(10):3913–3916. doi: 10.1073/pnas.71.10.3913. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Haberland M. E., Reynolds J. A. Interaction of L-alpha-palmitoyl lysophosphatidylcholine with the AI polypeptide of high density lipoprotein. J Biol Chem. 1975 Sep 10;250(17):6636–6639. [PubMed] [Google Scholar]
- Helenius A., Simons K. The binding of detergents to lipophilic and hydrophilic proteins. J Biol Chem. 1972 Jun 10;247(11):3656–3661. [PubMed] [Google Scholar]
- Le Maire M., Moller J. V., Tanford C. Retention of enzyme activity by detergent-solubilized sarcoplasmic Ca2+ -ATPase. Biochemistry. 1976 Jun 1;15(11):2336–2342. doi: 10.1021/bi00656a014. [DOI] [PubMed] [Google Scholar]
- Makino S., Reynolds J. A., Tanford C. The binding of deoxycholate and Triton X-100 to proteins. J Biol Chem. 1973 Jul 25;248(14):4926–4932. [PubMed] [Google Scholar]
- Makino S., Tanford C., Reynolds J. A. The interaction of polypeptide components of human high density serum lipoprotein with detergents. J Biol Chem. 1974 Dec 10;249(23):7379–7382. [PubMed] [Google Scholar]
- Reynolds J. A., Simon R. H. The interaction of polypeptide components of human high density lipoprotein with sodium dodecyl sulfate. J Biol Chem. 1974 Jun 25;249(12):3937–3940. [PubMed] [Google Scholar]
- Sardet C., Tardieu A., Luzzati V. Shape and size of bovine rhodopsin: a small-angle x-ray scattering study of a rhodopsin-detergent complex. J Mol Biol. 1976 Aug 15;105(3):383–407. doi: 10.1016/0022-2836(76)90100-5. [DOI] [PubMed] [Google Scholar]
- Schoenborn B. P. Neutron scattering for the analysis of membranes. Biochim Biophys Acta. 1976 Apr 13;457(1):41–55. doi: 10.1016/0304-4157(76)90013-7. [DOI] [PubMed] [Google Scholar]
- Siegel L. M., Monty K. J. Determination of molecular weights and frictional ratios of proteins in impure systems by use of gel filtration and density gradient centrifugation. Application to crude preparations of sulfite and hydroxylamine reductases. Biochim Biophys Acta. 1966 Feb 7;112(2):346–362. doi: 10.1016/0926-6585(66)90333-5. [DOI] [PubMed] [Google Scholar]
- Smith R., Dawson J. R., Tanford C. The size and number of polypeptide chains in human serum low density lipoprotein. J Biol Chem. 1972 Jun 10;247(11):3376–3381. [PubMed] [Google Scholar]
- Tanford C., Nozaki Y., Reynolds J. A., Makino S. Molecular characterization of proteins in detergent solutions. Biochemistry. 1974 May 21;13(11):2369–2376. doi: 10.1021/bi00708a021. [DOI] [PubMed] [Google Scholar]
- Tausk R. J., van Esch J., Karmiggelt J., Voordouw G., Overbeek J. T. Physical chemical studies of short-chain lecithin homologues. II. Micellar weights of dihexanoyl- and diheptanoyllecithin. Biophys Chem. 1974 Feb;1(3):184–203. doi: 10.1016/0301-4622(74)80005-0. [DOI] [PubMed] [Google Scholar]
- Tomita M., Marchesi V. T. Amino-acid sequence and oligosaccharide attachment sites of human erythrocyte glycophorin. Proc Natl Acad Sci U S A. 1975 Aug;72(8):2964–2968. doi: 10.1073/pnas.72.8.2964. [DOI] [PMC free article] [PubMed] [Google Scholar]
- VINOGRAD J., HEARST J. E. Equilibrium sedimentation of macromolecules and viruses in a density gradient. Fortschr Chem Org Naturst. 1962;20:373–422. [PubMed] [Google Scholar]