Protein Stretching III: Force-Extension Curves of Tethered Bovine Carbonic Anhydrase B to the Silicon Substrate under Native, Intermediate and Denaturing Conditions
- 1 June 1999
- journal article
- Published by IOP Publishing in Japanese Journal of Applied Physics
- Vol. 38 (6S) , 3912
- https://doi.org/10.1143/jjap.38.3912
Abstract
Atomic force microscope (AFM) was used to measure the force-extension relationship of the globular protein, carbonic anhydrase B, having 259 amino acid residues, under native, denaturing and intermediate solution conditions. For this purpose, the protein was genetically engineered in order to endow it with -SH groups at its N- and C- termini and was fixed to the silanized surface of a silicon wafer using a covalent cross-linker with the reactive end to -SH and with a long spacer of polyethylene glycol. The silicon nitride tip of the AFM was covered with a cross-linker with the reactive end to -SH and was brought into contact with the protein on the silicon surface. Subsequent force curve measurements showed occasional stable bond formation, and the force extension relationship obtained from such curves showed distinct characteristics of the native, denatured and intermediate forms of carbonic anhydrase B.Keywords
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