Modern analytical ultracentrifugation in protein science: A tutorial review
By J. Lebowitz, M. Lewis, Peter Schuck. Protein Science, 2002
755 citations recorded by the source corpus.
Analytical ultracentrifugation (AU) is reemerging as a versatile tool for the study of proteins. Monitoring the sedimentation of macromolecules in the centrifugal field allows their hydrodynamic and thermodynamic characterization in solution, without interaction with any matrix or surface. The combination of new instrumentation and powerful computational software for data analysis has led to major advances in the characterization of proteins and protein complexes. The pace of new advancements makes it difficult for protein scientists to gain sufficient expertise to apply modern AU to their research problems. To address this problem, this review builds from the basic concepts to advanced approaches for the characterization of protein systems, and key computational and internet resources are provided. We will first explore the characterization of proteins by sedimentation velocity (SV). Determination of sedimentation coefficients allows for the modeling of the hydrodynamic shape of proteins and protein complexes. The computational treatment of SV data to resolve sedimenting components has been achieved. Hence, SV can be very useful in the identification of the oligomeric state and the stoichiometry of heterogeneous interactions. The second major part of the review covers sedimentation equilibrium (SE) of proteins, including membrane proteins and glycoproteins. This is the method of choice for molar mass determinations and the study of self‐association and heterogeneous interactions, such as protein–protein, protein–nucleic acid, and protein–small molecule binding.
Subfields: Materials Chemistry, Molecular Biology
View Paper: https://www.semanticscholar.org/paper/07975bba7a9f710c3ce23d90953d180faa323d60
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Citations in the corpus
- A de novo designed protein–protein interface (2007, methodology): “Data were processed using the WinNONLIN software from the National Analytical Ultracentrifuge Facility following standard global nonlinear fitting methods (Lebowitz et al. 2002).”
- Average protein density is a molecular‐weight‐dependent function (2004, methodology): “Later on, Tsai et al. (1999) performed theoretical calculations using the average volume of buried residues to evaluate the proteins’ density and compare them with experimental data corresponding to 12 proteins taken from the compilation by Squire and Himmel (1979) and Gekko and Noguchi (1979).”
- Sedimentation equilibrium analysis of protein interactions with global implicit mass conservation constraints and systematic noise decomposition. (2004, background): “Recent reviews, and introductory texts on principle and practice can be found in (3-12).”
- Challenges in the development of high protein concentration formulations. (2004, background): “The impact of hydrodynamic volume leading to apparently larger proteins than actual size is especially noted for highly glycosylated proteins that have shapes different from the typical globular protein standards (Shire 1992; Lebowitz et al. 2002 ).”
- Protein aggregation: pathways, induction factors and analysis. (2009, methodology): “This method was pioneered by Svedberg already in the early 1940s,(145) however due to the advanced development of instrumentation technology and computational software nowadays AUC has become a broadly utilized tool in the analysis of proteins.(140) There are a number of reviews on”
- On the analysis of protein self-association by sedimentation velocity analytical ultracentrifugation. (2003, methodology): “(8-12)), largely due to the ability to study reversible interactions in solution and the increasing interest in protein interactions.”
- Electrophoretic mobility shift assay (EMSA) for detecting protein–nucleic acid interactions (2007, methodology): “and investigators are also encouraged to consider non-EMSA methods such as analytical ultracentrifugatio”