Protein tryptophan accessibility studied by fluorescence quenching

Abstract

A laboratory class is described to introduce the biochemistry major students to the basic concepts and various applications of fluorescence spectroscopy. Through simple and inexpensive experiments the students learn how to record excitation and emission spectra and measure intrinsic protein fluorescence and its quenching to elucidate the local tryptophan environment. Free tryptophan, ovalbumin, and bovine serum albumin are used for the experiments. The structural information that can be retrieved from emission spectra, as well as from quenching data, is discussed. © 2002 by The International Union of Biochemistry and Molecular Biology.