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Using High-performance Liquid Chromatography to Measure the Effects of Protein-stabilizing Cosolvents on a Model Protein and Fluorescent Probes by Michael Lever; Madhusudan K. Vasudevamurthy; Marie A. Squire is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

What is Using High-performance Liquid Chromatography to Measure the Effects of Protein-stabilizing Cosolvents on a Model Protein and Fluorescent Probes about?

The effect of cosolvents on the fluorescence of solutes was measured manually and in an automated high-performance liquid chromatography (HPLC) system that eliminates fluorescent contaminants on-line. The HPLC system was used to show that the effect of cosolvents on the fluorescence spectrum of heated chymotrypsin (a measure of unfolding) correlates with the effect of the solutes on the heat stabilization of catalytic activity; r 2 = 0.73 with 12 example cosolvents. Changes in the fluorescence of model probes showed that known counteracting solutes slightly decrease the polarity of the solvent. Different cosolvents affect the proton transfer indicator, 2-naphthol (a model for tyrosinyl residues) differently, polyhydric alcohols enhance the protonated naphthol emission whereas zwitterionic solutes enhance naphthoxide fluorescence. The results with the automated system are consistent with the known stabilizing effects of the cosolvents and validate it as a tool to explore the development of novel cosolvents and their effects on multiple biological systems.

Who reads Using High-performance Liquid Chromatography to Measure the Effects of Protein-stabilizing Cosolvents on a Model Protein and Fluorescent Probes?

Author

Michael Lever; Madhusudan K. Vasudevamurthy; Marie A. Squire

Publisher

Elsevier Science; Elsevier ; Elsevier Inc.; Elsevier BV (ISSN 0003-2697)

Published

2007

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)