Article
Dissociation kinetics of an enzyme-inhibitor system using single-molecule force measurements
Biomacromolecules
(2010)
Abstract
We report on an improved method to interpret single molecule dissociation measurements using atomic force microscopy. We describe an easy to use methodology to reject nonspecific binding events, as well as estimating the number of multiple binding events. The method takes nonlinearities in the force profiles into account that result from the deformation of the used polymeric linkers. This new method is applied to a relevant enzyme−inhibitor system, latent matrix metalloprotease 9 (ProMMP-9, a gelatinase), and its inhibitor, tissue inhibitor of metalloproteases 1 (TIMP 1), which are important players in cancer metastasis. Our method provides a measured kinetic off-rate of 0.010 ± 0.003 s−1 for the dissociation of ProMMP9 and TIMP1, which is consistent with values measured by ensemble methods.
Disciplines
Publication Date
2010
DOI
https://doi.org/10.1021/bm100844x
Citation Information
Peter M. Hoffmann, Essa Mayyas, Margarida Bernardo, Lindsay Runyan, et al.. "Dissociation kinetics of an enzyme-inhibitor system using single-molecule force measurements" Biomacromolecules (2010) Available at: http://works.bepress.com/peter_m_hoffmann/18/