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Resonance energy transfer between green fluorescent protein variants: complexities revealed with myosin fusion proteins
Authors:Zeng Wei  Seward Harriet E  Málnási-Csizmadia András  Wakelin Stuart  Woolley Robert J  Cheema Gurpreet S  Basran Jaswir  Patel Trushar R  Rowe Arthur J  Bagshaw Clive R
Institution:Department of Biochemistry, University of Leicester, UK.
Abstract:Green fluorescent protein and its variants are frequently used as F?rster (fluorescence) resonance energy transfer (FRET) pairs to determine the proximity of protein domains. We prepared fusion proteins comprising yellow fluorescent protein-Dictyostelium myosin II motor domain-cyan fluorescent protein (YFP-myosin-CFP) and compared their FRET properties with an existing construct (GFP-myosin-BFP), containing a green fluorescent protein acceptor and blue fluorescent protein donor Suzuki, Y., Yasunaga, T., Ohkura, R., Wakabayashi, T. and Sutoh, K. (1998) Nature 396, 380-383]. The latter construct showed an apparent 40% reduction in acceptor fluorescence on ATP addition, when excited via the donor, compared with the YFP-myosin-CFP constructs which showed a small increase (
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