Broos, Jaap,

Gonelli, Margherita,

Robillard, George T.,

Schuurman-Wolters, Gea,

Strambini, Giovanni B.,

Vos, Erwin P.P.,

Broos, Jaap

Strambini, Giovanni B.

Gonelli, Margherita

Schuurman-Wolters, Gea

Robillard, George T.

University of Groningen Research Database

  
 University of Groningen
Studying Membrane Transport Protein Dynamics with Tryptophan Phosphorescence
Spectroscopy
Broos, Jaap; Strambini, Giovanni B.; Gonelli, Margherita; Schuurman-Wolters, Gea; Robillard,
George T.
IMPORTANT NOTE: You are advised to consult the publisher's version (publisher's PDF) if you wish to cite from
it. Please check the document version below.
Document Version
Publisher's PDF, also known as Version of record
Publication date:
2001
Link to publication in University of Groningen/UMCG research database
Citation for published version (APA):
Broos, J., Strambini, G. B., Gonelli, M., Schuurman-Wolters, G., & Robillard, G. T., (2001). Studying
Membrane Transport Protein Dynamics with Tryptophan Phosphorescence Spectroscopy, 1 p.
Copyright
Other than for strictly personal use, it is not permitted to download or to forward/distribute the text or part of it without the consent of the
author(s) and/or copyright holder(s), unless the work is under an open content license (like Creative Commons).
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Download date: 10-02-2018
STUDYING MEMBRANE TRANSPORT PROTEIN DYNAMICS WITH 
TRYPTOPHAN PHOSPHORESCENCE SPECROSCOPY 
Jaap Broos1, Giovanni B. Strambini2, Margherita Gonelli2, Gea Schuurman Wolters1, 
Erwin P.P. Vos1, George T. Robillard1: 1University of Groningen, Biochemistry, 
Nyenborgh 4, Groningen, 9747AG Netherlands, 2CNR, biofisica, via Alfieri 1, Pisa, 
56010 Italy 
A detailed description of the mechanism of membrane-bound transporters requires 
the characterization of the conformations of the protein involved in the different stages of 
solute translocation. However these changes are either are not detected by conventional 
biophysical methods or, as in the case of fluorescence, are not readily ascribed to specific 
structural alterations. In the past decade, the phosphorescence emission of tryptophan 
residues was shown to provide an exquisitely sensitive monitor of the local protein 
structure. We implemented this technique for membrane proteins as demonstrated for the 
mannitol transporter, EII mtl , from E. coli. This protein is responsible for the transport of 
mannitol across the inner membrane and its concomitant phosphorylation to mannitol 1-
phosphate. Phosphorescence decays in buffer revealed large variations of the triplet 
lifetimes of the wild-type protein and six single-tryptophan-containing mutants. Mannitol 
binding induces a more ordered- and homogeneous structure near the mannitol binding 
site. In contrast, enzyme phosphorylation induces a large relaxation of the protein 
structure at the reporter sites. The implications of these structural changes on the coupling 
mechanism between the transport and the phosphorylation activity of EII mtl are 
discussed. 
 


Studying Membrane Transport Protein Dynamics with Tryptophan Phosphorescence Spectroscopy

NARCIS 

ARTS repository - University of Groningen

   University of GroningenStudying Membrane Transport Protein Dynamics with Tryptophan PhosphorescenceSpectroscopyBroos, Jaap; Strambini, Giovanni B.; Gonelli, Margherita; Schuurman-Wolters, Gea; Robillard,George T.IMPORTANT NOTE: You are advised to consult the publisher's version (publisher's PDF) if you wish to cite fromit. Please check the document version below.Document VersionPublisher's PDF, also known as Version of recordPublication date:2001Link to publication in University of Groningen/UMCG research databaseCitation for published version (APA):Broos, J., Strambini, G. B., Gonelli, M., Schuurman-Wolters, G., & Robillard, G. T. (2001). StudyingMembrane Transport Protein Dynamics with Tryptophan Phosphorescence Spectroscopy.CopyrightOther than for strictly personal use, it is not permitted to download or to forward/distribute the text or part of it without the consent of theauthor(s) and/or copyright holder(s), unless the work is under an open content license (like Creative Commons).The publication may also be distributed here under the terms of Article 25fa of the Dutch Copyright Act, indicated by the “Taverne” license.More information can be found on the University of Groningen website: https://www.rug.nl/library/open-access/self-archiving-pure/taverne-amendment.Take-down policyIf you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediatelyand investigate your claim.Downloaded from the University of Groningen/UMCG research database (Pure): http://www.rug.nl/research/portal. For technical reasons thenumber of authors shown on this cover page is limited to 10 maximum.Download date: 31-10-2023STUDYING MEMBRANE TRANSPORT PROTEIN DYNAMICS WITH TRYPTOPHAN PHOSPHORESCENCE SPECROSCOPY Jaap Broos1, Giovanni B. Strambini2, Margherita Gonelli2, Gea Schuurman Wolters1, Erwin P.P. Vos1, George T. Robillard1: 1University of Groningen, Biochemistry, Nyenborgh 4, Groningen, 9747AG Netherlands, 2CNR, biofisica, via Alfieri 1, Pisa, 56010 Italy A detailed description of the mechanism of membrane-bound transporters requires the characterization of the conformations of the protein involved in the different stages of solute translocation. However these changes are either are not detected by conventional biophysical methods or, as in the case of fluorescence, are not readily ascribed to specific structural alterations. In the past decade, the phosphorescence emission of tryptophan residues was shown to provide an exquisitely sensitive monitor of the local protein structure. We implemented this technique for membrane proteins as demonstrated for the mannitol transporter, EII mtl , from E. coli. This protein is responsible for the transport of mannitol across the inner membrane and its concomitant phosphorylation to mannitol 1-phosphate. Phosphorescence decays in buffer revealed large variations of the triplet lifetimes of the wild-type protein and six single-tryptophan-containing mutants. Mannitol binding induces a more ordered- and homogeneous structure near the mannitol binding site. In contrast, enzyme phosphorylation induces a large relaxation of the protein structure at the reporter sites. The implications of these structural changes on the coupling mechanism between the transport and the phosphorylation activity of EII mtl are discussed.  

English

University of Groningen Digital Archive

Proceedings - University of Groningen

   University of GroningenStudying Membrane Transport Protein Dynamics with Tryptophan PhosphorescenceSpectroscopyBroos, Jaap; Strambini, Giovanni B.; Gonelli, Margherita; Schuurman-Wolters, Gea; Robillard,George T.IMPORTANT NOTE: You are advised to consult the publisher's version (publisher's PDF) if you wish to cite fromit. Please check the document version below.Document VersionPublisher's PDF, also known as Version of recordPublication date:2001Link to publication in University of Groningen/UMCG research databaseCitation for published version (APA):Broos, J., Strambini, G. B., Gonelli, M., Schuurman-Wolters, G., & Robillard, G. T. (2001). StudyingMembrane Transport Protein Dynamics with Tryptophan Phosphorescence Spectroscopy.CopyrightOther than for strictly personal use, it is not permitted to download or to forward/distribute the text or part of it without the consent of theauthor(s) and/or copyright holder(s), unless the work is under an open content license (like Creative Commons).The publication may also be distributed here under the terms of Article 25fa of the Dutch Copyright Act, indicated by the “Taverne” license.More information can be found on the University of Groningen website: https://www.rug.nl/library/open-access/self-archiving-pure/taverne-amendment.Take-down policyIf you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediatelyand investigate your claim.Downloaded from the University of Groningen/UMCG research database (Pure): http://www.rug.nl/research/portal. For technical reasons thenumber of authors shown on this cover page is limited to 10 maximum.Download date: 11-09-2023STUDYING MEMBRANE TRANSPORT PROTEIN DYNAMICS WITH TRYPTOPHAN PHOSPHORESCENCE SPECROSCOPY Jaap Broos1, Giovanni B. Strambini2, Margherita Gonelli2, Gea Schuurman Wolters1, Erwin P.P. Vos1, George T. Robillard1: 1University of Groningen, Biochemistry, Nyenborgh 4, Groningen, 9747AG Netherlands, 2CNR, biofisica, via Alfieri 1, Pisa, 56010 Italy A detailed description of the mechanism of membrane-bound transporters requires the characterization of the conformations of the protein involved in the different stages of solute translocation. However these changes are either are not detected by conventional biophysical methods or, as in the case of fluorescence, are not readily ascribed to specific structural alterations. In the past decade, the phosphorescence emission of tryptophan residues was shown to provide an exquisitely sensitive monitor of the local protein structure. We implemented this technique for membrane proteins as demonstrated for the mannitol transporter, EII mtl , from E. coli. This protein is responsible for the transport of mannitol across the inner membrane and its concomitant phosphorylation to mannitol 1-phosphate. Phosphorescence decays in buffer revealed large variations of the triplet lifetimes of the wild-type protein and six single-tryptophan-containing mutants. Mannitol binding induces a more ordered- and homogeneous structure near the mannitol binding site. In contrast, enzyme phosphorylation induces a large relaxation of the protein structure at the reporter sites. The implications of these structural changes on the coupling mechanism between the transport and the phosphorylation activity of EII mtl are discussed.  

Dissertations of the University of Groningen

   University of GroningenStudying Membrane Transport Protein Dynamics with Tryptophan PhosphorescenceSpectroscopyBroos, Jaap; Strambini, Giovanni B.; Gonelli, Margherita; Schuurman-Wolters, Gea; Robillard,George T.IMPORTANT NOTE: You are advised to consult the publisher's version (publisher's PDF) if you wish to cite fromit. Please check the document version below.Document VersionPublisher's PDF, also known as Version of recordPublication date:2001Link to publication in University of Groningen/UMCG research databaseCitation for published version (APA):Broos, J., Strambini, G. B., Gonelli, M., Schuurman-Wolters, G., & Robillard, G. T. (2001). StudyingMembrane Transport Protein Dynamics with Tryptophan Phosphorescence Spectroscopy.CopyrightOther than for strictly personal use, it is not permitted to download or to forward/distribute the text or part of it without the consent of theauthor(s) and/or copyright holder(s), unless the work is under an open content license (like Creative Commons).The publication may also be distributed here under the terms of Article 25fa of the Dutch Copyright Act, indicated by the “Taverne” license.More information can be found on the University of Groningen website: https://www.rug.nl/library/open-access/self-archiving-pure/taverne-amendment.Take-down policyIf you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediatelyand investigate your claim.Downloaded from the University of Groningen/UMCG research database (Pure): http://www.rug.nl/research/portal. For technical reasons thenumber of authors shown on this cover page is limited to 10 maximum.Download date: 05-06-2022STUDYING MEMBRANE TRANSPORT PROTEIN DYNAMICS WITH TRYPTOPHAN PHOSPHORESCENCE SPECROSCOPY Jaap Broos1, Giovanni B. Strambini2, Margherita Gonelli2, Gea Schuurman Wolters1, Erwin P.P. Vos1, George T. Robillard1: 1University of Groningen, Biochemistry, Nyenborgh 4, Groningen, 9747AG Netherlands, 2CNR, biofisica, via Alfieri 1, Pisa, 56010 Italy A detailed description of the mechanism of membrane-bound transporters requires the characterization of the conformations of the protein involved in the different stages of solute translocation. However these changes are either are not detected by conventional biophysical methods or, as in the case of fluorescence, are not readily ascribed to specific structural alterations. In the past decade, the phosphorescence emission of tryptophan residues was shown to provide an exquisitely sensitive monitor of the local protein structure. We implemented this technique for membrane proteins as demonstrated for the mannitol transporter, EII mtl , from E. coli. This protein is responsible for the transport of mannitol across the inner membrane and its concomitant phosphorylation to mannitol 1-phosphate. Phosphorescence decays in buffer revealed large variations of the triplet lifetimes of the wild-type protein and six single-tryptophan-containing mutants. Mannitol binding induces a more ordered- and homogeneous structure near the mannitol binding site. In contrast, enzyme phosphorylation induces a large relaxation of the protein structure at the reporter sites. The implications of these structural changes on the coupling mechanism between the transport and the phosphorylation activity of EII mtl are discussed.  

   University of GroningenStudying Membrane Transport Protein Dynamics with Tryptophan PhosphorescenceSpectroscopyBroos, Jaap; Strambini, Giovanni B.; Gonelli, Margherita; Schuurman-Wolters, Gea; Robillard,George T.IMPORTANT NOTE: You are advised to consult the publisher's version (publisher's PDF) if you wish to cite fromit. Please check the document version below.Document VersionPublisher's PDF, also known as Version of recordPublication date:2001Link to publication in University of Groningen/UMCG research databaseCitation for published version (APA):Broos, J., Strambini, G. B., Gonelli, M., Schuurman-Wolters, G., & Robillard, G. T. (2001). StudyingMembrane Transport Protein Dynamics with Tryptophan Phosphorescence Spectroscopy.CopyrightOther than for strictly personal use, it is not permitted to download or to forward/distribute the text or part of it without the consent of theauthor(s) and/or copyright holder(s), unless the work is under an open content license (like Creative Commons).The publication may also be distributed here under the terms of Article 25fa of the Dutch Copyright Act, indicated by the “Taverne” license.More information can be found on the University of Groningen website: https://www.rug.nl/library/open-access/self-archiving-pure/taverne-amendment.Take-down policyIf you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediatelyand investigate your claim.Downloaded from the University of Groningen/UMCG research database (Pure): http://www.rug.nl/research/portal. For technical reasons thenumber of authors shown on this cover page is limited to 10 maximum.Download date: 04-06-2022STUDYING MEMBRANE TRANSPORT PROTEIN DYNAMICS WITH TRYPTOPHAN PHOSPHORESCENCE SPECROSCOPY Jaap Broos1, Giovanni B. Strambini2, Margherita Gonelli2, Gea Schuurman Wolters1, Erwin P.P. Vos1, George T. Robillard1: 1University of Groningen, Biochemistry, Nyenborgh 4, Groningen, 9747AG Netherlands, 2CNR, biofisica, via Alfieri 1, Pisa, 56010 Italy A detailed description of the mechanism of membrane-bound transporters requires the characterization of the conformations of the protein involved in the different stages of solute translocation. However these changes are either are not detected by conventional biophysical methods or, as in the case of fluorescence, are not readily ascribed to specific structural alterations. In the past decade, the phosphorescence emission of tryptophan residues was shown to provide an exquisitely sensitive monitor of the local protein structure. We implemented this technique for membrane proteins as demonstrated for the mannitol transporter, EII mtl , from E. coli. This protein is responsible for the transport of mannitol across the inner membrane and its concomitant phosphorylation to mannitol 1-phosphate. Phosphorescence decays in buffer revealed large variations of the triplet lifetimes of the wild-type protein and six single-tryptophan-containing mutants. Mannitol binding induces a more ordered- and homogeneous structure near the mannitol binding site. In contrast, enzyme phosphorylation induces a large relaxation of the protein structure at the reporter sites. The implications of these structural changes on the coupling mechanism between the transport and the phosphorylation activity of EII mtl are discussed.  

University of Groningen

   University of GroningenStudying Membrane Transport Protein Dynamics with Tryptophan PhosphorescenceSpectroscopyBroos, Jaap; Strambini, Giovanni B.; Gonelli, Margherita; Schuurman-Wolters, Gea; Robillard,George T.IMPORTANT NOTE: You are advised to consult the publisher's version (publisher's PDF) if you wish to cite fromit. Please check the document version below.Document VersionPublisher's PDF, also known as Version of recordPublication date:2001Link to publication in University of Groningen/UMCG research databaseCitation for published version (APA):Broos, J., Strambini, G. B., Gonelli, M., Schuurman-Wolters, G., & Robillard, G. T. (2001). StudyingMembrane Transport Protein Dynamics with Tryptophan Phosphorescence Spectroscopy.CopyrightOther than for strictly personal use, it is not permitted to download or to forward/distribute the text or part of it without the consent of theauthor(s) and/or copyright holder(s), unless the work is under an open content license (like Creative Commons).Take-down policyIf you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediatelyand investigate your claim.Downloaded from the University of Groningen/UMCG research database (Pure): http://www.rug.nl/research/portal. For technical reasons thenumber of authors shown on this cover page is limited to 10 maximum.Download date: 20-05-2020STUDYING MEMBRANE TRANSPORT PROTEIN DYNAMICS WITH TRYPTOPHAN PHOSPHORESCENCE SPECROSCOPY Jaap Broos1, Giovanni B. Strambini2, Margherita Gonelli2, Gea Schuurman Wolters1, Erwin P.P. Vos1, George T. Robillard1: 1University of Groningen, Biochemistry, Nyenborgh 4, Groningen, 9747AG Netherlands, 2CNR, biofisica, via Alfieri 1, Pisa, 56010 Italy A detailed description of the mechanism of membrane-bound transporters requires the characterization of the conformations of the protein involved in the different stages of solute translocation. However these changes are either are not detected by conventional biophysical methods or, as in the case of fluorescence, are not readily ascribed to specific structural alterations. In the past decade, the phosphorescence emission of tryptophan residues was shown to provide an exquisitely sensitive monitor of the local protein structure. We implemented this technique for membrane proteins as demonstrated for the mannitol transporter, EII mtl , from E. coli. This protein is responsible for the transport of mannitol across the inner membrane and its concomitant phosphorylation to mannitol 1-phosphate. Phosphorescence decays in buffer revealed large variations of the triplet lifetimes of the wild-type protein and six single-tryptophan-containing mutants. Mannitol binding induces a more ordered- and homogeneous structure near the mannitol binding site. In contrast, enzyme phosphorylation induces a large relaxation of the protein structure at the reporter sites. The implications of these structural changes on the coupling mechanism between the transport and the phosphorylation activity of EII mtl are discussed.  

   University of GroningenStudying Membrane Transport Protein Dynamics with Tryptophan PhosphorescenceSpectroscopyBroos, Jaap; Strambini, Giovanni B.; Gonelli, Margherita; Schuurman-Wolters, Gea; Robillard,George T.IMPORTANT NOTE: You are advised to consult the publisher's version (publisher's PDF) if you wish to cite fromit. Please check the document version below.Document VersionPublisher's PDF, also known as Version of recordPublication date:2001Link to publication in University of Groningen/UMCG research databaseCitation for published version (APA):Broos, J., Strambini, G. B., Gonelli, M., Schuurman-Wolters, G., & Robillard, G. T. (2001). StudyingMembrane Transport Protein Dynamics with Tryptophan Phosphorescence Spectroscopy.CopyrightOther than for strictly personal use, it is not permitted to download or to forward/distribute the text or part of it without the consent of theauthor(s) and/or copyright holder(s), unless the work is under an open content license (like Creative Commons).The publication may also be distributed here under the terms of Article 25fa of the Dutch Copyright Act, indicated by the “Taverne” license.More information can be found on the University of Groningen website: https://www.rug.nl/library/open-access/self-archiving-pure/taverne-amendment.Take-down policyIf you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediatelyand investigate your claim.Downloaded from the University of Groningen/UMCG research database (Pure): http://www.rug.nl/research/portal. For technical reasons thenumber of authors shown on this cover page is limited to 10 maximum.Download date: 01-11-2023STUDYING MEMBRANE TRANSPORT PROTEIN DYNAMICS WITH TRYPTOPHAN PHOSPHORESCENCE SPECROSCOPY Jaap Broos1, Giovanni B. Strambini2, Margherita Gonelli2, Gea Schuurman Wolters1, Erwin P.P. Vos1, George T. Robillard1: 1University of Groningen, Biochemistry, Nyenborgh 4, Groningen, 9747AG Netherlands, 2CNR, biofisica, via Alfieri 1, Pisa, 56010 Italy A detailed description of the mechanism of membrane-bound transporters requires the characterization of the conformations of the protein involved in the different stages of solute translocation. However these changes are either are not detected by conventional biophysical methods or, as in the case of fluorescence, are not readily ascribed to specific structural alterations. In the past decade, the phosphorescence emission of tryptophan residues was shown to provide an exquisitely sensitive monitor of the local protein structure. We implemented this technique for membrane proteins as demonstrated for the mannitol transporter, EII mtl , from E. coli. This protein is responsible for the transport of mannitol across the inner membrane and its concomitant phosphorylation to mannitol 1-phosphate. Phosphorescence decays in buffer revealed large variations of the triplet lifetimes of the wild-type protein and six single-tryptophan-containing mutants. Mannitol binding induces a more ordered- and homogeneous structure near the mannitol binding site. In contrast, enzyme phosphorylation induces a large relaxation of the protein structure at the reporter sites. The implications of these structural changes on the coupling mechanism between the transport and the phosphorylation activity of EII mtl are discussed.  

Studying Membrane Transport Protein Dynamics with Tryptophan Phosphorescence Spectroscopy

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University of Groningen Research Database

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Proceedings - University of Groningen

Dissertations of the University of Groningen

Proceedings - University of Groningen

Dissertations of the University of Groningen

University of Groningen

ARTS repository - University of Groningen