TY - JOUR
T1 - Overview of current methods in sedimentation velocity and sedimentation equilibrium analytical ultracentrifugation
AU - Zhao, Huaying
AU - Brautigam, Chad A
AU - Ghirlando, Rodolfo
AU - Schuck, Peter
PY - 2013/2
Y1 - 2013/2
N2 - Modern computational strategies have allowed for the direct modeling of the sedimentation process of heterogeneous mixtures, resulting in sedimentation velocity (SV) sizedistribution analyses with significantly improved detection limits and strongly enhanced resolution. These advances have transformed the practice of SV, rendering it the primary method of choice for most existing applications of analytical ultracentrifugation (AUC), such as the study of protein self- and hetero-association, the study of membrane proteins, and applications in biotechnology. New global multisignal modeling and mass conservation approaches in SV and sedimentation equilibrium (SE), in conjunction with the effective-particle framework for interpreting the sedimentation boundary structure of interacting systems, as well as tools for explicit modeling of the reaction/ diffusion/sedimentation equations to experimental data, have led to more robust and more powerful strategies for the study of reversible protein interactions and multiprotein complexes. Furthermore, modern mathematical modeling capabilities have allowed for a detailed description of many experimental aspects of the acquired data, thus enabling novel experimental opportunities, with important implications for both sample preparation and data acquisition. The goal of the current unit is to describe the current tools for the study of soluble proteins, detergent-solubilized membrane proteins and their interactions by SV and SE.
AB - Modern computational strategies have allowed for the direct modeling of the sedimentation process of heterogeneous mixtures, resulting in sedimentation velocity (SV) sizedistribution analyses with significantly improved detection limits and strongly enhanced resolution. These advances have transformed the practice of SV, rendering it the primary method of choice for most existing applications of analytical ultracentrifugation (AUC), such as the study of protein self- and hetero-association, the study of membrane proteins, and applications in biotechnology. New global multisignal modeling and mass conservation approaches in SV and sedimentation equilibrium (SE), in conjunction with the effective-particle framework for interpreting the sedimentation boundary structure of interacting systems, as well as tools for explicit modeling of the reaction/ diffusion/sedimentation equations to experimental data, have led to more robust and more powerful strategies for the study of reversible protein interactions and multiprotein complexes. Furthermore, modern mathematical modeling capabilities have allowed for a detailed description of many experimental aspects of the acquired data, thus enabling novel experimental opportunities, with important implications for both sample preparation and data acquisition. The goal of the current unit is to describe the current tools for the study of soluble proteins, detergent-solubilized membrane proteins and their interactions by SV and SE.
KW - Analytical ultracentrifugation
KW - Chemical equilibria
KW - Multiprotein complex
KW - Protein hydrodynamics
KW - Reversible interaction
KW - Sedimentation equilibrium
KW - Sedimentation velocity
KW - Size-distribution
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U2 - 10.1002/0471140864.ps2012s71
DO - 10.1002/0471140864.ps2012s71
M3 - Article
C2 - 23377850
AN - SCOPUS:84879936868
SN - 1934-3655
JO - Current Protocols in Protein Science
JF - Current Protocols in Protein Science
IS - SUPPL.71
M1 - 20.12
ER -