title: Detecting and exploring partially unfolded states of proteins using a sensor with chaperone bound to its surface.
abstract: We have developed a sensor concept capable of discriminating environments that induce proteins to enter unfolding intermediate states. Such a sensor detects the presence of environmental stressors such as chemical agents in aqueous media, thermal stress or the presence of ionizing or non-ionizing radiation by monitoring the conformation state of a "sensor protein". In this paper, we demonstrate the concept by using surface plasmon resonance to monitor binding of thermally and chemically stressed sensor proteins to a chaperone, alpha-crystallin, bound to the sensor surface. Citrate synthase and insulin were used as example sensor proteins to detect the presence of thermal stress and chemical stress, respectively. It was shown that alpha-crystallin retained its chaperone action after immobilization on the Biacore sensor chip. The binding of early and late unfolding intermediates of citrate synthase was discriminated using the association and dissociation behaviour of the binding. The sensor is therefore capable of assessing the severity of an environmental stress.
authors.affiliation: School of Physics, University of Sydney, NSW 2006, Australia.
authors.forename: Doaa F
authors.initials: DF
authors.lastname: George
authors.forename: Marcela M M
authors.initials: MM
authors.lastname: Bilek
authors.forename: David R
authors.initials: DR
authors.lastname: McKenzie
mesh_terms: D001665:Binding Sites; D015374:Biosensing Techniques / Q000379:methods*; D002950:Citrate (si)-Synthase / Q000737:chemistry* / Q000648:ultrastructure*; D003461:Crystallography / Q000379:methods*; D007328:Insulin / Q000737:chemistry*; D018832:Molecular Chaperones / Q000737:chemistry*; D011485:Protein Binding; D011489:Protein Denaturation; D017510:Protein Folding; D020349:Surface Plasmon Resonance / Q000379:methods*; D013499:Surface Properties; D013696:Temperature
chemical_list: D007328:Insulin; D018832:Molecular Chaperones; D002950:Citrate (si)-Synthase