Chapter title |
Using Light Scattering to Determine the Stoichiometry of Protein Complexes
|
---|---|
Chapter number | 14 |
Book title |
Protein-Protein Interactions
|
Published in |
Methods in molecular biology, January 2015
|
DOI | 10.1007/978-1-4939-2425-7_14 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2424-0, 978-1-4939-2425-7
|
Authors |
Jeremy Mogridge, Mogridge, Jeremy |
Abstract |
The stoichiometry of a protein complex can be calculated from an accurate measurement of the complex's molecular weight. Multiangle laser light scattering in combination with size exclusion chromatography and interferometric refractometry provides a powerful means for determining the molecular weights of proteins and protein complexes. In contrast to conventional size exclusion chromatography and analytical centrifugation, measurements do not rely on the use of molecular weight standards and are not affected by the shape of the proteins. The technique is based on the direct relationship between the amount of light scattered by a protein in solution, and the product of its concentration and molecular weight. A typical experimental configuration includes a size exclusion column to fractionate the sample, a light scattering detector to measure scattered light, and an interferometric refractometer to measure protein concentration. The determination of the molecular weight of an anthrax toxin complex will be used to illustrate how multiangle laser light scattering can be used to determine the stoichiometry of protein complexes. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
United States | 1 | 2% |
Netherlands | 1 | 2% |
Sweden | 1 | 2% |
Germany | 1 | 2% |
Unknown | 39 | 91% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Researcher | 16 | 37% |
Student > Ph. D. Student | 5 | 12% |
Student > Master | 4 | 9% |
Student > Bachelor | 3 | 7% |
Student > Doctoral Student | 2 | 5% |
Other | 7 | 16% |
Unknown | 6 | 14% |
Readers by discipline | Count | As % |
---|---|---|
Agricultural and Biological Sciences | 17 | 40% |
Biochemistry, Genetics and Molecular Biology | 10 | 23% |
Physics and Astronomy | 3 | 7% |
Chemistry | 3 | 7% |
Pharmacology, Toxicology and Pharmaceutical Science | 1 | 2% |
Other | 2 | 5% |
Unknown | 7 | 16% |