Abstract:
:The interaction of hen egg white lysozyme with the trisaccharide tri-N-acetyl glucosamine has been well-characterized by biophysical methods and structural biology. In this chapter, we present a series of experiments designed to detect and quantify that interaction using several commonly available biophysical methods: thermal shift assay, fluorescence intensity, microscale thermophoresis, isothermal titration calorimetry, and surface plasmon resonance.These experiments have been used for teaching and troubleshooting in a core facility. By taking a set of representative data from several years of practical courses, we are able to demonstrate the robustness of the protocols, calculate confidence intervals for the dissociation constant from each method, and illustrate the degree of consistency between those methods when applied to a simple system in a single location by different experimenters.
journal_name
Methods Mol Bioljournal_title
Methods in molecular biology (Clifton, N.J.)authors
Li-Blatter X,Zweifel L,Sharpe Tdoi
10.1007/978-1-0716-1197-5_2keywords:
["Dissociation constant","Fluorescence","Global fitting","Isothermal titration calorimetry","Microscale thermophoresis","Orthogonal assay","Surface plasmon resonance","Teaching","Thermal shift"]subject
Has Abstractpub_date
2021-01-01 00:00:00pages
47-79eissn
1064-3745issn
1940-6029journal_volume
2263pub_type
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