Tryptophan mutants of human C5a anaphylatoxin: A fluorescence anisotropy decay and energy transfer study

M. Federwisch; A. Wollmer; M. Emde; T. Stühmer; T. Melcher; A. Klos; J. Köhl; W. Bautsch

doi:10.1016/0301-4622(93)80017-D

Tryptophan mutants of human C5a anaphylatoxin: A fluorescence anisotropy decay and energy transfer study

M. Federwisch, A. Wollmer^*, M. Emde, T. Stühmer, T. Melcher, A. Klos, J. Köhl, W. Bautsch

^*Korrespondierende/r Autor/-in für diese Arbeit

Institut für Systemische Entzündungsforschung

11 Zitate (Scopus)

Abstract

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.

Originalsprache	Englisch
Zeitschrift	Biophysical Chemistry
Jahrgang	46
Ausgabenummer	3
Seiten (von - bis)	237-248
Seitenumfang	12
ISSN	0301-4622
DOIs	https://doi.org/10.1016/0301-4622(93)80017-D
Publikationsstatus	Veröffentlicht - 01.05.1993

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title = "Tryptophan mutants of human C5a anaphylatoxin: A fluorescence anisotropy decay and energy transfer study",

abstract = "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.",

author = "M. Federwisch and A. Wollmer and M. Emde and T. St{\"u}hmer and T. Melcher and A. Klos and J. K{\"o}hl and W. Bautsch",

year = "1993",

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doi = "10.1016/0301-4622(93)80017-D",

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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.

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U2 - 10.1016/0301-4622(93)80017-D

DO - 10.1016/0301-4622(93)80017-D

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C2 - 8343570

AN - SCOPUS:0027212582

SN - 0301-4622

VL - 46

SP - 237

EP - 248

JO - Biophysical Chemistry

JF - Biophysical Chemistry

IS - 3

ER -

Tryptophan mutants of human C5a anaphylatoxin: A fluorescence anisotropy decay and energy transfer study

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