Modified Rhodopsins From Aureobasidium pullulans Excel With Very High Proton-Transport Rates
Zitieren Sie bitte immer diese URN: urn:nbn:de:bvb:20-opus-249248
- Aureobasidium pullulans is a black fungus that can adapt to various stressful conditions like hypersaline, acidic, and alkaline environments. The genome of A. pullulans exhibits three genes coding for putative opsins ApOps1, ApOps2, and ApOps3. We heterologously expressed these genes in mammalian cells and Xenopus oocytes. Localization in the plasma membrane was greatly improved by introducing additional membrane trafficking signals at the N-terminus and the C-terminus. In patch-clamp and two-electrode-voltage clamp experiments, all threeAureobasidium pullulans is a black fungus that can adapt to various stressful conditions like hypersaline, acidic, and alkaline environments. The genome of A. pullulans exhibits three genes coding for putative opsins ApOps1, ApOps2, and ApOps3. We heterologously expressed these genes in mammalian cells and Xenopus oocytes. Localization in the plasma membrane was greatly improved by introducing additional membrane trafficking signals at the N-terminus and the C-terminus. In patch-clamp and two-electrode-voltage clamp experiments, all three proteins showed proton pump activity with maximal activity in green light. Among them, ApOps2 exhibited the most pronounced proton pump activity with current amplitudes occasionally extending 10 pA/pF at 0 mV. Proton pump activity was further supported in the presence of extracellular weak organic acids. Furthermore, we used site-directed mutagenesis to reshape protein functions and thereby implemented light-gated proton channels. We discuss the difference to other well-known proton pumps and the potential of these rhodopsins for optogenetic applications.…
Autor(en): | Sabine Panzer, Chong Zhang, Tilen Konte, Celine Bräuer, Anne Diemar, Parathy Yogendran, Jing Yu-Strzelczyk, Georg Nagel, Shiqiang Gao, Ulrich Terpitz |
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URN: | urn:nbn:de:bvb:20-opus-249248 |
Dokumentart: | Artikel / Aufsatz in einer Zeitschrift |
Institute der Universität: | Medizinische Fakultät / Physiologisches Institut |
Fakultät für Biologie / Theodor-Boveri-Institut für Biowissenschaften | |
Sprache der Veröffentlichung: | Englisch |
Titel des übergeordneten Werkes / der Zeitschrift (Englisch): | Frontiers in Molecular Biosciences |
ISSN: | 2296-889X |
Erscheinungsjahr: | 2021 |
Band / Jahrgang: | 8 |
Aufsatznummer: | 750528 |
Originalveröffentlichung / Quelle: | Frontiers in Molecular Biosciences (2021) 8:750528. doi: 10.3389/fmolb.2021.750528 |
DOI: | https://doi.org/10.3389/fmolb.2021.750528 |
Allgemeine fachliche Zuordnung (DDC-Klassifikation): | 6 Technik, Medizin, angewandte Wissenschaften / 61 Medizin und Gesundheit / 610 Medizin und Gesundheit |
Freie Schlagwort(e): | Aureobasidium; black yeast; fungal rhodopsins; membrane trafficking; microbial rhodopsins; optogenetics; photoreceptor; proton channel |
Datum der Freischaltung: | 11.02.2022 |
Datum der Erstveröffentlichung: | 01.11.2021 |
Open-Access-Publikationsfonds / Förderzeitraum 2021 | |
Lizenz (Deutsch): | CC BY: Creative-Commons-Lizenz: Namensnennung 4.0 International |