| Publication type: | Article in scientific journal |
| Type of review: | Peer review (publication) |
| Title: | The citrate carrier CitS probed by single-molecule fluorescence spectroscopy |
| Authors: | Kastner, C. Prummer, M. Sick, Beate Renn, A. Wild, U.P. Dimroth, P. |
| DOI: | 10.1016/S0006-3495(03)74974-1 |
| Published in: | Biophysical Journal |
| Volume(Issue): | 84 |
| Issue: | 3 |
| Page(s): | 1651 |
| Pages to: | 1659 |
| Issue Date: | 2003 |
| Publisher / Ed. Institution: | Cell Press |
| ISSN: | 1542-0086 0006-3495 |
| Language: | English |
| Subjects: | Optics; Membrane protein; Single molecule; Biology |
| Subject (DDC): | 572: Biochemistry |
| Abstract: | A prominent region of the Naþ-dependent citrate carrier (CitS) from Klebsiella pneumoniae is the highly conserved loop X-XI, which contains a putative citrate binding site. To monitor potential conformational changes within this region by single-molecule fluorescence spectroscopy, the target cysteines C398 and C414 of the single-Cys mutants (CitSsC398, CitS-sC414) were selectively labelled with the thiol-reactive fluorophores AlexaFluor 546/568 C5 maleimide (AF546, AF568). While both single-cysteine mutants were catalytically active citrate carriers, labelling with the fluorophore was only tolerated at C398. Upon citrate addition to the functional protein fluorophore conjugate CitS-sC398-AF546, complete fluorescence quenching of the majority of molecules was observed, indicating a citrate-induced conformational change of the fluorophore-containing domain of CitS. This quenching was specific for the physiological substrate citrate and therefore most likely reflecting a conformational change in the citrate transport mechanism. Single-molecule studies with dual-labelled CitSsC398-AF546/568 and dual-color detection provided strong evidence for a homodimeric association of CitS. |
| Further description: | Content available via Elsevier Open Archive |
| URI: | https://digitalcollection.zhaw.ch/handle/11475/14038 |
| Fulltext version: | Published version |
| License (according to publishing contract): | Licence according to publishing contract |
| Departement: | School of Engineering |
| Organisational Unit: | Institute of Data Analysis and Process Design (IDP) |
| Appears in collections: | Publikationen School of Engineering |
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Kastner, C., Prummer, M., Sick, B., Renn, A., Wild, U. P., & Dimroth, P. (2003). The citrate carrier CitS probed by single-molecule fluorescence spectroscopy. Biophysical Journal, 84(3), 1651–1659. https://doi.org/10.1016/S0006-3495(03)74974-1
Kastner, C. et al. (2003) ‘The citrate carrier CitS probed by single-molecule fluorescence spectroscopy’, Biophysical Journal, 84(3), pp. 1651–1659. Available at: https://doi.org/10.1016/S0006-3495(03)74974-1.
C. Kastner, M. Prummer, B. Sick, A. Renn, U. P. Wild, and P. Dimroth, “The citrate carrier CitS probed by single-molecule fluorescence spectroscopy,” Biophysical Journal, vol. 84, no. 3, pp. 1651–1659, 2003, doi: 10.1016/S0006-3495(03)74974-1.
KASTNER, C., M. PRUMMER, Beate SICK, A. RENN, U.P. WILD und P. DIMROTH, 2003. The citrate carrier CitS probed by single-molecule fluorescence spectroscopy. Biophysical Journal. 2003. Bd. 84, Nr. 3, S. 1651–1659. DOI 10.1016/S0006-3495(03)74974-1
Kastner, C., M. Prummer, Beate Sick, A. Renn, U.P. Wild, and P. Dimroth. 2003. “The Citrate Carrier CitS Probed by Single-Molecule Fluorescence Spectroscopy.” Biophysical Journal 84 (3): 1651–59. https://doi.org/10.1016/S0006-3495(03)74974-1.
Kastner, C., et al. “The Citrate Carrier CitS Probed by Single-Molecule Fluorescence Spectroscopy.” Biophysical Journal, vol. 84, no. 3, 2003, pp. 1651–59, https://doi.org/10.1016/S0006-3495(03)74974-1.
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