Abstract
It has now become increasingly clear that a complete atomic description of how biomacromolecules recognize each other requires knowledge not only of the structures of the complexes but also of how kinetics and thermodynamics drive the binding process. In particular, such knowledge is lacking for protein–glycosaminoglycan (GAG) complexes. Isothermal titration calorimetry (ITC) is the only technique that can provide various thermodynamic parameters—enthalpy, entropy, free energy (binding constant), and stoichiometry—from a single experiment. Here we describe different factors that must be taken into consideration in carrying out ITC titrations to obtain meaningful thermodynamic data of protein–GAG interactions.
An erratum to this chapter is available at http://dx.doi.org/10.1007/978-1-4939-1714-3_48
An erratum to this chapter can be found at http://dx.doi.org/10.1007/978-1-4939-1714-3_48
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References
Lindahl U, Kjellén L (2013) Pathophysiology of heparan sulphate: many diseases, few drugs. J Intern Med 273:555–571
Bishop JR, Schuksz M, Esko JD (2007) Heparan sulphate proteoglycans fine-tune mammalian physiology. Nature 446:1030–1037
Ladbury JE, Klebe G, Freire E (2010) Adding calorimetric data to decision making in lead discovery: a hot tip. Nat Rev Drug Discov 9:23–27
Salanga CL, Handel TM (2011) Chemokine oligomerization and interactions with receptors and glycosaminoglycans: the role of structural dynamics in function. Exp Cell Res 317:590–601
Gangavarapu P, Rajagopalan L, Kolli D, Guerrero-Plata A, Garofalo RP, Rajarathnam K (2012) The monomer-dimer equilibrium and glycosaminoglycan interactions of chemokine CXCL8 regulate tissue-specific neutrophil recruitment. J Leukoc Biol 91:259–265
Kuschert GSV, Coulin F, Power CA, Proudfoot AEI, Hubbard RE, Hoogewerf AJ, Wells TNC (1999) Glycosaminoglycans interact selectively with chemokines and modulate receptor binding and cellular responses. Biochemistry 38:12959–12968
Pace CN, Vajdos F, Fee L, Grimsley G, Gray T (1995) How to measure and predict the molar absorption coefficient of a protein. Protein Sci 4:2411–2423
Rajarathnam K, Rösgen J (2013) Isothermal titration calorimetry of membrane proteins – progress and challenges. Biochim Biophys Acta (in press). doi: pii: S0005-2736(13)00169-7. 10.1016/j.bbamem.2013.05.023
Venkataraman G, Sasisekharan V, Herr AB, Ornitz DM, Waksman G, Cooney CL, Langer R, Sasisekharan R (1996) Preferential self-association of basic fibroblast growth factor is stabilized by heparin during receptor dimerization and activation. Proc Natl Acad Sci 93:845–850
Wiseman T, Williston S, Brandts JF, Lin L-N (1989) Rapid measurement of binding constants and heats of binding using a new titration calorimeter. Anal Biochem 179:131–137
Acknowledgements
This work was supported in part by grants P01HL1071521 and R21AI097975 to K.R. and R01GM049760 to J.R. from the National Institutes of Health. The authors would like to thank Dr. Luis Holthauzen for technical support.
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Dutta, A.K., Rösgen, J., Rajarathnam, K. (2015). Using Isothermal Titration Calorimetry to Determine Thermodynamic Parameters of Protein–Glycosaminoglycan Interactions. In: Balagurunathan, K., Nakato, H., Desai, U. (eds) Glycosaminoglycans. Methods in Molecular Biology, vol 1229. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1714-3_25
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DOI: https://doi.org/10.1007/978-1-4939-1714-3_25
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