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Fluorescent labelling of proteins - Volume 1

Chappell, D. C. (1991). Fluorescent labelling of proteins - Volume 1. (Unpublished Doctoral thesis, City, University of London)

Abstract

A range of fluorescent probes were utilised for the fluorescent modification, via both amine and thiol groups, of a protein that binds cAMP. The protein concerned, called cAMP Receptor Protein, undergoes a conformational change on cAMP binding, and as a consequence, the microenvironment of the probe is altered with regard to polarity. The probes were then screened in order to ascertain the degree of sensitivity to the polarity of their environment. This property was monitored via alterations in the fluorescence characteristics of the probe, particularly fluorescence enhancement and/or shifts in peak emission. The most suitable probes may then be applied to a protein that binds cGMP, and the alteration in the fluorescence characteristics induced in the probe via cGMP binding, may then be utilised in order to determine the intracellular cGMP concentration within the intact cell.

Publication Type: Thesis (Doctoral)
Subjects: Q Science > QD Chemistry
Departments: Doctoral Theses
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