Unfolding Individual Domains of BmrA, a Bacterial ABC Transporter Involved in Multidrug Resistance
- Publikationstyp:
- Zeitschriftenaufsatz
- Metadaten:
-
- Autoren
- Kristin Oepen
- Veronika Mater
- Dirk Schneider
- Autoren-URL
- https://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=fis-test-1&SrcAuth=WosAPI&KeyUT=WOS:000959188200001&DestLinkType=FullRecord&DestApp=WOS_CPL
- DOI
- 10.3390/ijms24065239
- eISSN
- 1422-0067
- Externe Identifier
- Clarivate Analytics Document Solution ID: C0VA5
- PubMed Identifier: 36982314
- Ausgabe der Veröffentlichung
- 6
- Zeitschrift
- INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
- Schlüsselwörter
- ABC transporter
- BmrA
- membrane protein
- multi-domain protein
- protein folding
- protein stability
- Trp fluorescence spectroscopy
- Artikelnummer
- ARTN 5239
- Datum der Veröffentlichung
- 2023
- Status
- Published
- Titel
- Unfolding Individual Domains of BmrA, a Bacterial ABC Transporter Involved in Multidrug Resistance
- Sub types
- Article
- Ausgabe der Zeitschrift
- 24
Datenquelle: Web of Science (Lite)
- Andere Metadatenquellen:
-
- Abstract
- <jats:p>The folding and stability of proteins are often studied via unfolding (and refolding) a protein with urea. Yet, in the case of membrane integral protein domains, which are shielded by a membrane or a membrane mimetic, urea generally does not induce unfolding. However, the unfolding of α-helical membrane proteins may be induced by the addition of sodium dodecyl sulfate (SDS). When protein unfolding is followed via monitoring changes in Trp fluorescence characteristics, the contributions of individual Trp residues often cannot be disentangled, and, consequently, the folding and stability of the individual domains of a multi-domain membrane protein cannot be studied. In this study, the unfolding of the homodimeric bacterial ATP-binding cassette (ABC) transporter Bacillus multidrug resistance ATP (BmrA), which comprises a transmembrane domain and a cytosolic nucleotide-binding domain, was investigated. To study the stability of individual BmrA domains in the context of the full-length protein, the individual domains were silenced by mutating the existent Trps. The SDS-induced unfolding of the corresponding constructs was compared to the (un)folding characteristics of the wild-type (wt) protein and isolated domains. The full-length variants BmrAW413Y and BmrAW104YW164A were able to mirror the changes observed with the isolated domains; thus, these variants allowed for the study of the unfolding and thermodynamic stability of mutated domains in the context of full-length BmrA.</jats:p>
- Autoren
- Kristin Oepen
- Veronika Mater
- Dirk Schneider
- DOI
- 10.3390/ijms24065239
- eISSN
- 1422-0067
- Ausgabe der Veröffentlichung
- 6
- Zeitschrift
- International Journal of Molecular Sciences
- Sprache
- en
- Online publication date
- 2023
- Paginierung
- 5239 - 5239
- Status
- Published online
- Herausgeber
- MDPI AG
- Herausgeber URL
- http://dx.doi.org/10.3390/ijms24065239
- Datum der Datenerfassung
- 2023
- Titel
- Unfolding Individual Domains of BmrA, a Bacterial ABC Transporter Involved in Multidrug Resistance
- Ausgabe der Zeitschrift
- 24
Datenquelle: Crossref
- Abstract
- The folding and stability of proteins are often studied via unfolding (and refolding) a protein with urea. Yet, in the case of membrane integral protein domains, which are shielded by a membrane or a membrane mimetic, urea generally does not induce unfolding. However, the unfolding of α-helical membrane proteins may be induced by the addition of sodium dodecyl sulfate (SDS). When protein unfolding is followed via monitoring changes in Trp fluorescence characteristics, the contributions of individual Trp residues often cannot be disentangled, and, consequently, the folding and stability of the individual domains of a multi-domain membrane protein cannot be studied. In this study, the unfolding of the homodimeric bacterial ATP-binding cassette (ABC) transporter <i>Bacillus</i> multidrug resistance ATP (BmrA), which comprises a transmembrane domain and a cytosolic nucleotide-binding domain, was investigated. To study the stability of individual BmrA domains in the context of the full-length protein, the individual domains were silenced by mutating the existent Trps. The SDS-induced unfolding of the corresponding constructs was compared to the (un)folding characteristics of the wild-type (wt) protein and isolated domains. The full-length variants BmrA<sub>W413Y</sub> and BmrA<sub>W104YW164A</sub> were able to mirror the changes observed with the isolated domains; thus, these variants allowed for the study of the unfolding and thermodynamic stability of mutated domains in the context of full-length BmrA.
- Addresses
- Department of Chemistry-Biochemistry, Johannes Gutenberg University Mainz, Hanns-Dieter-Hüsch-Weg 17, 55128 Mainz, Germany.
- Autoren
- Kristin Oepen
- Veronika Mater
- Dirk Schneider
- DOI
- 10.3390/ijms24065239
- eISSN
- 1422-0067
- Externe Identifier
- PubMed Identifier: 36982314
- PubMed Central ID: PMC10049088
- Funding acknowledgements
- DynaMem (State of Rhineland-Palatinate): N/A
- Open access
- true
- ISSN
- 1422-0067
- Ausgabe der Veröffentlichung
- 6
- Zeitschrift
- International journal of molecular sciences
- Schlüsselwörter
- Bacillus
- Urea
- ATP-Binding Cassette Transporters
- Adenosine Triphosphate
- Drug Resistance, Multiple, Bacterial
- Protein Folding
- Protein Unfolding
- Sprache
- eng
- Medium
- Electronic
- Online publication date
- 2023
- Open access status
- Open Access
- Paginierung
- 5239
- Datum der Veröffentlichung
- 2023
- Status
- Published
- Publisher licence
- CC BY
- Datum der Datenerfassung
- 2023
- Titel
- Unfolding Individual Domains of BmrA, a Bacterial ABC Transporter Involved in Multidrug Resistance.
- Sub types
- research-article
- Journal Article
- Ausgabe der Zeitschrift
- 24
Files
https://europepmc.org/articles/PMC10049088?pdf=render
Datenquelle: Europe PubMed Central
- Abstract
- The folding and stability of proteins are often studied via unfolding (and refolding) a protein with urea. Yet, in the case of membrane integral protein domains, which are shielded by a membrane or a membrane mimetic, urea generally does not induce unfolding. However, the unfolding of α-helical membrane proteins may be induced by the addition of sodium dodecyl sulfate (SDS). When protein unfolding is followed via monitoring changes in Trp fluorescence characteristics, the contributions of individual Trp residues often cannot be disentangled, and, consequently, the folding and stability of the individual domains of a multi-domain membrane protein cannot be studied. In this study, the unfolding of the homodimeric bacterial ATP-binding cassette (ABC) transporter Bacillus multidrug resistance ATP (BmrA), which comprises a transmembrane domain and a cytosolic nucleotide-binding domain, was investigated. To study the stability of individual BmrA domains in the context of the full-length protein, the individual domains were silenced by mutating the existent Trps. The SDS-induced unfolding of the corresponding constructs was compared to the (un)folding characteristics of the wild-type (wt) protein and isolated domains. The full-length variants BmrAW413Y and BmrAW104YW164A were able to mirror the changes observed with the isolated domains; thus, these variants allowed for the study of the unfolding and thermodynamic stability of mutated domains in the context of full-length BmrA.
- Date of acceptance
- 2023
- Autoren
- Kristin Oepen
- Veronika Mater
- Dirk Schneider
- Autoren-URL
- https://www.ncbi.nlm.nih.gov/pubmed/36982314
- DOI
- 10.3390/ijms24065239
- eISSN
- 1422-0067
- Externe Identifier
- PubMed Central ID: PMC10049088
- Funding acknowledgements
- DynaMem (State of Rhineland-Palatinate): N/A
- Ausgabe der Veröffentlichung
- 6
- Zeitschrift
- Int J Mol Sci
- Schlüsselwörter
- ABC transporter
- BmrA
- Trp fluorescence spectroscopy
- membrane protein
- multi-domain protein
- protein folding
- protein stability
- Adenosine Triphosphate
- ATP-Binding Cassette Transporters
- Protein Folding
- Protein Unfolding
- Urea
- Drug Resistance, Multiple, Bacterial
- Bacillus
- Sprache
- eng
- Country
- Switzerland
- PII
- ijms24065239
- Datum der Veröffentlichung
- 2023
- Status
- Published online
- Datum, an dem der Datensatz öffentlich gemacht wurde
- 2023
- Titel
- Unfolding Individual Domains of BmrA, a Bacterial ABC Transporter Involved in Multidrug Resistance.
- Sub types
- Journal Article
- Ausgabe der Zeitschrift
- 24
Datenquelle: PubMed
- Beziehungen:
- Eigentum von