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Proceedings Paper

Frequency-domain fluorescence of mutant cytochrome b5
Author(s): Alexey S. Ladokhin; Henryk M. Malak; Michael L. Johnson; Joseph R. Lakowicz; L. Wang; A. W. Steggles; Peter W. Holloway
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Paper Abstract

Cytochrome b5, isolated by detergent extraction from rabbit liver, has been extensively studied by fluorescence techniques, however, its fluorescence properties are complicated by the presence of three tryptophans in the membrane-binding domain. This protein has now been expressed in E. coli and a mutant form has been isolated which contains only one tryptophan (Trp-109) in the membrane-binding domain. The mutant protein does differ from the native protein in several of its spectroscopic properties, but it interacts with lipids in a similar way and shows the same functional activity. The availability of a mutant cytochrome b5 with a single Trp in the nonpolar domain enables several frequency-domain experiments: to estimate the distribution of distances between Trp-109 and the heme; to study the incorporation of the protein into the membrane; to study the quenching of the tryptophan fluorescence by lipids with a brominated acyl chain; and to analyze the shape of the lifetime distribution.

Paper Details

Date Published: 1 April 1992
PDF: 8 pages
Proc. SPIE 1640, Time-Resolved Laser Spectroscopy in Biochemistry III, (1 April 1992); doi: 10.1117/12.58250
Show Author Affiliations
Alexey S. Ladokhin, Univ. of Virginia School of Medicine (United States)
Henryk M. Malak, Univ. of Maryland School of Medicine (United States)
Michael L. Johnson, Univ. of Virginia School of Medicine (United States)
Joseph R. Lakowicz, Univ. of Maryland School of Medicine (United States)
L. Wang, Northeastern Ohio Univ. College of Medicine (United States)
A. W. Steggles, Northeastern Ohio Univ. College of Medicine (United States)
Peter W. Holloway, Univ. of Virginia School of Medicine (United States)


Published in SPIE Proceedings Vol. 1640:
Time-Resolved Laser Spectroscopy in Biochemistry III
Joseph R. Lakowicz, Editor(s)

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