TY - JOUR
T1 - Signal and binding. I. Physico-chemical response to macromolecule–ligand interactions
AU - Bujalowski, Wlodzimierz
AU - Jezewska, Maria J.
AU - Bujalowski, Paul J.
N1 - Publisher Copyright:
© 2016
PY - 2017/3/1
Y1 - 2017/3/1
N2 - Obtaining a detailed knowledge about energetics of ligand–macromolecule interactions is a prerequisite for elucidation of the nature, behavior, and activities of the formed complexes. The most commonly used methods in characterizing molecular interactions are physico-chemical techniques based mainly on spectroscopic, calorimetric, hydrodynamic, etc., measurements. The major advantage of the physico-chemical methods is that they do not require large quantities of material and, if performed carefully, do not perturb examined reactions. Applications of several different physico-chemical approaches, commonly encountered in analyses of biochemical interactions, are here reviewed and discussed, using examples of simple binding reactions. It is stressed that without determination of the relationship between the measured signal and the total average degree of binding, the performed analysis of a single physico-chemical titration curve may provide only fitting parameters, instead of meaningful interaction parameters, already for the binding systems with only two ligand molecules. Some possible pitfalls in the analyses of single titration curves are discussed.
AB - Obtaining a detailed knowledge about energetics of ligand–macromolecule interactions is a prerequisite for elucidation of the nature, behavior, and activities of the formed complexes. The most commonly used methods in characterizing molecular interactions are physico-chemical techniques based mainly on spectroscopic, calorimetric, hydrodynamic, etc., measurements. The major advantage of the physico-chemical methods is that they do not require large quantities of material and, if performed carefully, do not perturb examined reactions. Applications of several different physico-chemical approaches, commonly encountered in analyses of biochemical interactions, are here reviewed and discussed, using examples of simple binding reactions. It is stressed that without determination of the relationship between the measured signal and the total average degree of binding, the performed analysis of a single physico-chemical titration curve may provide only fitting parameters, instead of meaningful interaction parameters, already for the binding systems with only two ligand molecules. Some possible pitfalls in the analyses of single titration curves are discussed.
KW - Binding isotherms
KW - Physico-chemical titrations
KW - Thermodynamics
UR - http://www.scopus.com/inward/record.url?scp=85009198115&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85009198115&partnerID=8YFLogxK
U2 - 10.1016/j.bpc.2016.12.006
DO - 10.1016/j.bpc.2016.12.006
M3 - Review article
C2 - 28092802
AN - SCOPUS:85009198115
SN - 0301-4622
VL - 222
SP - 7
EP - 24
JO - Biophysical Chemistry
JF - Biophysical Chemistry
ER -