Studying Macromolecular Interactions of Cellular Machines by the Combined Use of Analytical Ultracentrifugation, Light Scattering, and Fluorescence Spectroscopy Methods

Adv Exp Med Biol. 2024;3234:89-107. doi: 10.1007/978-3-031-52193-5_7.ABSTRACTCellular machines formed by the interaction and assembly of macromolecules are essential in many processes of the living cell. These assemblies involve homo- and hetero-associations, including protein-protein, protein-DNA, protein-RNA, and protein-polysaccharide associations, most of which are reversible. This chapter describes the use of analytical ultracentrifugation, light scattering, and fluorescence-based methods, well-established biophysical techniques, to characterize interactions leading to the formation of macromolecular complexes and their modulation in response to specific or unspecific factors. We also illustrate, with several examples taken from studies on bacterial processes, the advantages of the combined use of subsets of these techniques as orthogonal analytical methods to analyze protein oligomerization and polymerization, interactions with ligands, hetero-associations involving membrane proteins, and protein-nucleic acid complexes.PMID:38507202 | DOI:10.1007/978-3-031-52193-5_7

Source: Advances in Experimental Medicine and Biology - March 20, 2024 Category: Research Authors: Carlos Alfonso Marta Sobrinos-Sanguino Juan Rom án Luque-Ortega Silvia Zorrilla Bego ña Monterroso Oscar M Nuero Germ án Rivas Source Type: research