Fluorescence‐quenching‐resolved spectroscopy of proteins
A new procedure is described for using fluorescence‐quenching data of tryptophan residues in proteins to resolve their fluorescence emission spectra. In this concept the Stern‐Volmer quenching plot is determined at each particular emission wavelength and itterative non‐linear least‐squares fitting p...
Published in: | European Journal of Biochemistry |
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1988
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crwiley:10.1111/j.1432-1033.1988.tb13948.x 2024-06-02T08:14:53+00:00 Fluorescence‐quenching‐resolved spectroscopy of proteins WASYLEWSKI, Zygmunt KOLOCZEK, Henryk WASNIOWSKA, Alicja 1988 http://dx.doi.org/10.1111/j.1432-1033.1988.tb13948.x https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1111%2Fj.1432-1033.1988.tb13948.x https://febs.onlinelibrary.wiley.com/doi/pdf/10.1111/j.1432-1033.1988.tb13948.x en eng Wiley http://onlinelibrary.wiley.com/termsAndConditions#vor European Journal of Biochemistry volume 172, issue 3, page 719-724 ISSN 0014-2956 1432-1033 journal-article 1988 crwiley https://doi.org/10.1111/j.1432-1033.1988.tb13948.x 2024-05-03T10:44:37Z A new procedure is described for using fluorescence‐quenching data of tryptophan residues in proteins to resolve their fluorescence emission spectra. In this concept the Stern‐Volmer quenching plot is determined at each particular emission wavelength and itterative non‐linear least‐squares fitting procedure allowed to resolve the steady‐state emission spectra into components. The resolved components, attributed to each of tryptophan residue, can be characterized by different accessibility to the quencher. The ability to resolve fluorescence emission spectra can be improved by using different kinds of efficient quenchers, which can selectively quench the emission of exposed or both exposed and buried fluorophores. The method was used to decompose emission fluorescence spectra in two‐tryptophan‐containing proteins; horse liver dehydrogenase, sperm whale apomyoglobin and metalloprotease from Staphylococcus aureus. The resolved spectra of alcohol dehydrogenase and metalloprotease are in excellent agreement with those previously obtained by single‐photon counting or phase methods. The method presented here is technically simple and does not require expensive instrumentation. Article in Journal/Newspaper Sperm whale Wiley Online Library European Journal of Biochemistry 172 3 719 724 |
institution |
Open Polar |
collection |
Wiley Online Library |
op_collection_id |
crwiley |
language |
English |
description |
A new procedure is described for using fluorescence‐quenching data of tryptophan residues in proteins to resolve their fluorescence emission spectra. In this concept the Stern‐Volmer quenching plot is determined at each particular emission wavelength and itterative non‐linear least‐squares fitting procedure allowed to resolve the steady‐state emission spectra into components. The resolved components, attributed to each of tryptophan residue, can be characterized by different accessibility to the quencher. The ability to resolve fluorescence emission spectra can be improved by using different kinds of efficient quenchers, which can selectively quench the emission of exposed or both exposed and buried fluorophores. The method was used to decompose emission fluorescence spectra in two‐tryptophan‐containing proteins; horse liver dehydrogenase, sperm whale apomyoglobin and metalloprotease from Staphylococcus aureus. The resolved spectra of alcohol dehydrogenase and metalloprotease are in excellent agreement with those previously obtained by single‐photon counting or phase methods. The method presented here is technically simple and does not require expensive instrumentation. |
format |
Article in Journal/Newspaper |
author |
WASYLEWSKI, Zygmunt KOLOCZEK, Henryk WASNIOWSKA, Alicja |
spellingShingle |
WASYLEWSKI, Zygmunt KOLOCZEK, Henryk WASNIOWSKA, Alicja Fluorescence‐quenching‐resolved spectroscopy of proteins |
author_facet |
WASYLEWSKI, Zygmunt KOLOCZEK, Henryk WASNIOWSKA, Alicja |
author_sort |
WASYLEWSKI, Zygmunt |
title |
Fluorescence‐quenching‐resolved spectroscopy of proteins |
title_short |
Fluorescence‐quenching‐resolved spectroscopy of proteins |
title_full |
Fluorescence‐quenching‐resolved spectroscopy of proteins |
title_fullStr |
Fluorescence‐quenching‐resolved spectroscopy of proteins |
title_full_unstemmed |
Fluorescence‐quenching‐resolved spectroscopy of proteins |
title_sort |
fluorescence‐quenching‐resolved spectroscopy of proteins |
publisher |
Wiley |
publishDate |
1988 |
url |
http://dx.doi.org/10.1111/j.1432-1033.1988.tb13948.x https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1111%2Fj.1432-1033.1988.tb13948.x https://febs.onlinelibrary.wiley.com/doi/pdf/10.1111/j.1432-1033.1988.tb13948.x |
genre |
Sperm whale |
genre_facet |
Sperm whale |
op_source |
European Journal of Biochemistry volume 172, issue 3, page 719-724 ISSN 0014-2956 1432-1033 |
op_rights |
http://onlinelibrary.wiley.com/termsAndConditions#vor |
op_doi |
https://doi.org/10.1111/j.1432-1033.1988.tb13948.x |
container_title |
European Journal of Biochemistry |
container_volume |
172 |
container_issue |
3 |
container_start_page |
719 |
op_container_end_page |
724 |
_version_ |
1800738889925656576 |