Pretransition and progressive softening of bovine carbonic anhydrase II as probed by single molecule atomic force microscopy
- 1 June 2005
- journal article
- Published by Wiley in Protein Science
- Vol. 14 (6), 1447-1457
- https://doi.org/10.1110/ps.041282305
Abstract
To develop a simple method for probing the physical state of surface adsorbed proteins, we adopted the force curve mode of an atomic force microscope (AFM) to extract information on the mechanical properties of surface immobilized bovine carbonic anhydrase II under native conditions and in the course of guanidinium chloride-induced denaturation. A progressive increase in the population of individually softened molecules was probed under mildly to fully denaturing conditions. The use of the approach regime of force curves gave information regarding the height and rigidity of the molecule under compressive stress, whereas use of the retracting regime of the curves gave information about the tensile characteristics of the protein. The results showed that protein molecules at the beginning of the transition region possessed slightly more flattened and significantly more softened conformations compared with that of native molecules, but were still not fully denatured, in agreement with results based on solution studies. Thus the force curve mode of an AFM was shown to be sensitive enough to provide information concerning the different physical states of single molecules of globular proteins.Keywords
This publication has 44 references indexed in Scilit:
- Unfolding a Folding Disease: Folding, Misfolding and Aggregation of the Marble Brain Syndrome-associated Mutant H107Y of Human Carbonic Anhydrase IIJournal of Molecular Biology, 2004
- Force Measurement and Inhibitor Binding Assay of Monomer and Engineered Dimer of Bovine Carbonic Anhydrase BBiochemical and Biophysical Research Communications, 2001
- Adsorption of Human Carbonic Anhydrase II Variants to Silica Nanoparticles Occur Stepwise: Binding Is Followed by Successive Conformational Changes to a Molten-Globule-like StateLangmuir, 2000
- Mechanical unfolding of a2‐macroglobulin molecules with atomic force microscopeFEBS Letters, 1996
- Direct Measurement for Elasticity of Myosin HeadBiochemical and Biophysical Research Communications, 1995
- Entropic Elasticity of λ-Phage DNAScience, 1994
- Structure of native and apo carbonic anhydrase II and structure of some of its anion-ligand complexesJournal of Molecular Biology, 1992
- On Compression of Rubber Elastic Sphere Over a Large Range of Displacements—Part 2: Comparison of Theory and ExperimentJournal of Engineering Materials and Technology, 1991
- On Compression of Rubber Elastic Sphere Over a Large Range of Displacements—Part 1: Theoretical StudyJournal of Engineering Materials and Technology, 1991
- Extensive Theory of Force-Approach Relations of Elastic Spheres in Compression and in ImpactJournal of Engineering Materials and Technology, 1989