title: Biophysical Characterisation and Quantification of Nucleic Acid-Protein Interactions: EMSA, MST and SPR.
abstract: Cell viability is only possible due to a dynamic range of essential nucleic acid-protein complex formation. DNA replication and repair, gene expression, transcription and protein synthesis are well-known processes mediated by nucleic acids (DNA and RNA) - protein interactions. Novel nucleic acid- protein complexes have been identified in the past few years aided by the development of numerous new techniques such as RNA capture or Tandem RNA Affinity Purification (TRAP). However, the biophysical and biochemical details of these interactions are mostly unknown. Here, we present three techniques (Electrophoretic Mobility Shift Assays, Microscale Thermophoresis and Surface Plasmon Resonance) that are commonly used to quantify and characterize DNA-protein and RNA-protein interactions and discuss their main advantages and limitations.
authors.forename: Janine K
authors.initials: JK
authors.lastname: Flores
authors.forename: Ruvini
authors.initials: R
authors.lastname: Kariawasam
authors.forename: Adrian X
authors.initials: AX
authors.lastname: Gimenez
authors.forename: Stephanie
authors.initials: S
authors.lastname: Helder
authors.forename: Liza
authors.initials: L
authors.lastname: Cubeddu
authors.forename: Roland
authors.initials: R
authors.lastname: Gamsjaeger
authors.affiliation: School of Molecular Bioscience, University of Sydney, NSW, 2006, Australia and School of Science and Health, University of Western Sydney, Penrith, NSW, 2751, Australia. sandro.ataide@sydney.edu.au.
authors.forename: Sandro F
authors.initials: SF
authors.lastname: Ataide
mesh_terms: D055592:Biophysical Phenomena; D024202:Electrophoretic Mobility Shift Assay / Q000379:methods*; D006801:Humans; D009696:Nucleic Acids / Q000378:metabolism*; D011485:Protein Binding; D012313:RNA / Q000378:metabolism; D018271:Signal Recognition Particle / Q000378:metabolism; D020349:Surface Plasmon Resonance; D062125:Thermometry / Q000379:methods*
chemical_list: D009696:Nucleic Acids; C503636:SRP72 protein, human; D018271:Signal Recognition Particle; D012313:RNA