TY - JOUR
T1 - Tryptophan mutants of human C5a anaphylatoxin: A fluorescence anisotropy decay and energy transfer study
AU - Federwisch, M.
AU - Wollmer, A.
AU - Emde, M.
AU - Stühmer, T.
AU - Melcher, T.
AU - Klos, A.
AU - Köhl, J.
AU - Bautsch, W.
PY - 1993/5/1
Y1 - 1993/5/1
N2 - Three mutants of the anaphylatoxin C5a were prepared with positions 2, 64 and 70, respectively, substituted by tryptophan. The last mutant was additionally labelled at Cys27 for fluorescence energy transfer (FET) measurements. The structural integrity and biological activity of the molecules were not affected. Fluorescence anisotropy decay (FAD) measurements showed that the rotational correlation time for tryptophan decreases in the order: [Trp2]rhC5a > [Trp64]rhC5a > [Trp70]rhC5a, indicating an increasing mobility of the side chain. Measurements of the fluorescence energy transfer from Trp70 to the 1,5-AEDANS group at Cys27 yielded a distance distribution of 2.4 ± 0.8 nm. This value is compatible with the C-terminal chain being arranged as a slightly stretched helix pointing away from the body of the molecule.
AB - Three mutants of the anaphylatoxin C5a were prepared with positions 2, 64 and 70, respectively, substituted by tryptophan. The last mutant was additionally labelled at Cys27 for fluorescence energy transfer (FET) measurements. The structural integrity and biological activity of the molecules were not affected. Fluorescence anisotropy decay (FAD) measurements showed that the rotational correlation time for tryptophan decreases in the order: [Trp2]rhC5a > [Trp64]rhC5a > [Trp70]rhC5a, indicating an increasing mobility of the side chain. Measurements of the fluorescence energy transfer from Trp70 to the 1,5-AEDANS group at Cys27 yielded a distance distribution of 2.4 ± 0.8 nm. This value is compatible with the C-terminal chain being arranged as a slightly stretched helix pointing away from the body of the molecule.
UR - http://www.scopus.com/inward/record.url?scp=0027212582&partnerID=8YFLogxK
U2 - 10.1016/0301-4622(93)80017-D
DO - 10.1016/0301-4622(93)80017-D
M3 - Journal articles
C2 - 8343570
AN - SCOPUS:0027212582
SN - 0301-4622
VL - 46
SP - 237
EP - 248
JO - Biophysical Chemistry
JF - Biophysical Chemistry
IS - 3
ER -
