Guide to Biophysical Methods for Macromolecular Characterization - 7

of dilution, a control experiment to measure heats of
binding is required.
ITC rapidly quantifies the energetics of non-covalent
binding between two proteins by analyzing
the heat released or absorbed resulting from the
proteins' conformational changes on binding, plus
the heat involved in the rearrangement of solvent
molecules near the binding site. ITC provides a
complete thermodynamic description of the interaction,
plus binding stoichiometry and the association/dissociation
constant. Additionally, when
structural information is available, ITC identifies the
contributions of specific amino acids involved and
their thermodynamic contributions.
Conclusion
Protein-protein interactions are involved in nearly
all physiological processes, including those associated
with health and disease. Discovery of these
complex, multi-factorial relationships has altered
the concept of target-based drug discovery and, in
fact, our understanding of basic biology. Quantifying
the thermodynamics of protein-protein interactions
typically involves the application of several
orthogonal analytic methods, including biochemical,
genetic, instrumental, and thermal-analytic
techniques. Two biophysical thermal analysis
methods, DSC and ITC, provide the widest depth
and breadth of characterization by quantifying all
relevant thermodynamics related to protein stability
and protein-protein binding.
References
1. Ijax, M., Ansari, MR, Iqbal, M. Intro Chapter: ProteinProtein
Interactions and Assays. July 18, 2018.
2. Protein Protein Interaction webpage. ScienceDirect.
2019.
3. Discovery and analysis of protein complexes.
Presentation. University of Alabama at Birmingham.
Targeted Metabolomics and Proteomics Laboratory.
4. Neduva, Victor and Russell, Robert B. (2005), Linear
motifs: Evolutionary interaction switches, FEBS Letters,
579, doi: 10.1016/j.febslet.2005.04.005
5. Nooren, I.M. and Thornton, J.M. (2003), Diversity of
protein-protein interactions. The EMBO Journal, 22:
3486-3492. https://doi.org/10.1093/emboj/cdg359
6. Shahar Sukenik, Pin Ren, Martin Gruebele. (2017)
Detection of weak protein complexes in live cells. PNAS,
114 (26) 6776-6781; DOI: 10.1073/pnas.1700818114
7. Schuck, P. and Braswell, E.H. (2000), Measuring ProteinProtein
Interactions by Equilibrium Sedimentation.
Current Protocols in Immunology, 40: 18.8.1-18.8.22.
https://doi.org/10.1002/0471142735.im1808s40
8. Corbett D., Bye J.W., Curtis R.A. (2019) Measuring
Nonspecific Protein-Protein Interactions by Dynamic
Light Scattering. In: McManus J. (eds) Protein SelfAssembly.
Methods in Molecular Biology, vol 2039.
https://doi.org/10.1007/978-1-4939-9678-0_1
9. Smits AH, Vermeulen M. (2016) Characterizing ProteinProtein
Interactions Using Mass Spectrometry:
Challenges and Opportunities. Trends Biotechnol.
Oct;34(10):825-834. doi: 10.1016/j.tibtech.2016.02.014.
10. K. H. Young, Yeast Two-hybrid: So Many Interactions, (in)
So Little Time..., Biology of Reproduction, Volume 58,
Issue 2, 1 February 1998, Pages 302-311, https://doi.
org/10.1095/biolreprod58.2.302
11. Lin JS, Lai EM. Protein-Protein Interactions: CoImmunoprecipitation.
Methods Mol Biol. 2017;1615:211219.
doi: 10.1007/978-1-4939-7033-9_17.
12. Sultana, A., and Lee, J.E. 2015. Measuring ProteinProtein
and Protein‐Nucleic Acid Interactions by
Biolayer Interferometry. Curr. Protoc. Protein Sci. 79:
19.25.1‐ 19.25.26. doi: 10.1002/0471140864.ps1925s79
13. Isothermal Titration Calorimetry webpage. Center for
Macromolecular Interactions. Harvard Medical School.
7

https://www.doi.org/10.1093/emboj/cdg359 https://www.doi.org/10.1002/0471142735.im1808s40 https://www.doi.org/10.1007/978-1-4939-9678-0_1 https://doi.org/10.1095/biolreprod58.2.302 https://doi.org/10.1095/biolreprod58.2.302

Guide to Biophysical Methods for Macromolecular Characterization

Table of Contents for the Digital Edition of Guide to Biophysical Methods for Macromolecular Characterization