Print Email Facebook Twitter Rapid and Highly Stable Membrane Reconstitution by LAiR Enables the Study of Physiological Integral Membrane Protein Functions Title Rapid and Highly Stable Membrane Reconstitution by LAiR Enables the Study of Physiological Integral Membrane Protein Functions Author Godoy Hernandez, A. (TU Delft BT/Biocatalysis) Asseri, Amer H. (King Abdulaziz University; Vrije Universiteit Amsterdam) Purugganan, Aiden J. (Student TU Delft) Jiko, Chimari (Institute for Integrated Radiation and Nuclear Science, Kyoto University) de Ram, C. Lill, Holger (Vrije Universiteit Amsterdam) Pabst, Martin (TU Delft BT/Environmental Biotechnology) Mitsuoka, Kaoru (Osaka University) Gerle, Christoph (Osaka University) Bald, Dirk (Vrije Universiteit Amsterdam) McMillan, D.G.G. (TU Delft BT/Biocatalysis; University of Tokyo) Date 2023 Abstract Functional reintegration into lipid environments represents a major challenge for in vitro investigation of integral membrane proteins (IMPs). Here, we report a new approach, termed LMNG Auto-insertion Reintegration (LAiR), for reintegration of IMPs into lipid bilayers within minutes. The resulting proteoliposomes displayed an unprecedented capability to maintain proton gradients and long-term stability. LAiR allowed for monitoring catalysis of a membrane-bound, physiologically relevant polyisoprenoid quinone substrate by Escherichia coli cytochromes bo3 (cbo3) and bd (cbd) under control of the proton motive force. LAiR also facilitated bulk-phase detection and physiological assessment of the “proton leak” in cbo3, a controversial catalytic state that previously was only approachable at the single-molecule level. LAiR maintained the multisubunit integrity and higher-order oligomeric states of the delicate mammalian F-ATP synthase. Given that LAiR can be applied to both liposomes and planar membrane bilayers and is compatible with IMPs and lipids from prokaryotic and eukaryotic sources, we anticipate LAiR to be applied broadly across basic research, pharmaceutical applications, and biotechnology. To reference this document use: http://resolver.tudelft.nl/uuid:f1bb28e5-929b-45c1-96ad-5896edaef509 DOI https://doi.org/10.1021/acscentsci.2c01170 ISSN 2374-7943 Source ACS Central Science, 9 (3), 494-507 Part of collection Institutional Repository Document type journal article Rights © 2023 A. Godoy Hernandez, Amer H. Asseri, Aiden J. Purugganan, Chimari Jiko, C. de Ram, Holger Lill, Martin Pabst, Kaoru Mitsuoka, Christoph Gerle, Dirk Bald, D.G.G. McMillan Files PDF acscentsci.2c01170.pdf 7.03 MB Close viewer /islandora/object/uuid:f1bb28e5-929b-45c1-96ad-5896edaef509/datastream/OBJ/view