Characterizing specific phage-protein interactions by fluorescence correlation spectroscopy

Protein Science : a Publication of the Protein Society
John T BahnsLiaohai Chen

Abstract

The interactions of several affinity reagent displayed T7 and M13 phage particles with their corresponding target molecules were examined using Fluorescence Correlation Spectroscopy (FCS). Diffusion times, relative fractions of each component in the recognition reactions at the equilibrium state, and ultimately the dissociation constants were deduced from analyzing the fluorescence autocorrelation curves. Although the sample preparation and FCS characterization of icosahedral T7-related systems were relatively straight forward, procedures with filamentous M13-related systems were complicated by the physical size of M13 and its aggregate formation. Methods that accommodate the FCS measurement of the M13 phage via changing confocal optics, fitting procedures, and aggregate discrimination are presented and discussed.

References

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Citations

Apr 11, 2017·Biochemistry·Dennis R GouletWilliam M Atkins
Aug 30, 2008·Journal of Biomolecular Screening·Helmi R M SchlamanHerman P Spaink
Aug 16, 2006·Analytical Biochemistry·Yuexing ZhangLiaohai Chen
Feb 16, 2011·Langmuir : the ACS Journal of Surfaces and Colloids·Nicholas A Kurniawan, Raj Rajagopalan

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