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Kinetics of Histidine-Tagged Protein Association to Nickel-Decorated Liposome Surfaces

Cite this: Langmuir 2019, 35, 38, 12550–12561
Publication Date (Web):August 29, 2019
https://doi.org/10.1021/acs.langmuir.9b01700
Copyright © 2019 American Chemical Society

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    Abstract

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    Nickel-chelating lipids offer a convenient platform for reversible immobilization of histidine-tagged proteins to liposome surfaces. This interaction recently found utility as a model system for studying membrane remodeling triggered by protein crowding. Despite its wide array of utility, the molecular details of transient protein association to the lipid surfaces decorated with such chelator lipids remains poorly understood. In this study, we explore the kinetics of protein–liposome association across a wide concentration range using stopped-flow fluorescence. The fluorescence of histidine-tagged protein containing an intrinsic fluorophore (superfolder green fluorescent protein, SfGFP) was quenched upon binding to Ni–NTA-modified liposomes containing the quencher Dabsyl-PE lipids. Stopped-flow fluorescence reveals a complex, multiexponential binding behavior with a fast (kobs ∼ 10–20 s–1) phase and slower (kobs < 4 s–1) phase. Interestingly, the observed rates for the slower phase increase initially under low concentrations but start decreasing once a critical concentration is reached. Despite differences in the binding time scales, we observe that the trend of decreasing rates is reproducible irrespective of the chelator lipid doping level, protein surface charge, or lipid composition. Consideration of the protein footprint and membrane surface area occupancy leads us to conclude that the multiphasic binding behavior is reflective of protein binding via two distinct binding conformations. We propose that preliminary steps in protein association involve binding of a sterically occlusive side-on conformation followed by reorganization that leads to an end-on conformation with increased packing density. These results are important for the improvement of histidine-tag-based immobilization strategies and offer mechanistic insight into intermediates preceding membrane bending driven by protein crowding.

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    The Supporting Information is available free of charge on the ACS Publications website at DOI: 10.1021/acs.langmuir.9b01700.

    • Materials and methods section and additional figures and tables (Figures S1–S15 and Tables S1 and S2) (PDF)

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    Cited By

    This article is cited by 11 publications.

    1. Yanjun Zheng, Tristan Wegner, Daniele Di Iorio, Marco Pierau, Frank Glorius, Seraphine V. Wegner. NTA-Cholesterol Analogue for the Nongenetic Liquid-Ordered Phase-Specific Functionalization of Lipid Membranes with Proteins. ACS Chemical Biology 2023, 18 (6) , 1435-1443. https://doi.org/10.1021/acschembio.3c00180
    2. Timothy Q. Vu, Lucas E. Sant’Anna, Neha P. Kamat. Tuning Targeted Liposome Avidity to Cells via Lipid Phase Separation. Biomacromolecules 2023, 24 (4) , 1574-1584. https://doi.org/10.1021/acs.biomac.2c01338
    3. Lucia Baldauf, Lennard van Buren, Federico Fanalista, Gijsje Hendrika Koenderink. Actomyosin-Driven Division of a Synthetic Cell. ACS Synthetic Biology 2022, 11 (10) , 3120-3133. https://doi.org/10.1021/acssynbio.2c00287
    4. Maryna Löwe, Sebastian Hänsch, Eymen Hachani, Lutz Schmitt, Stefanie Weidtkamp‐Peters, Alexej Kedrov. Probing macromolecular crowding at the lipid membrane interface with genetically‐encoded sensors. Protein Science 2023, 32 (11) https://doi.org/10.1002/pro.4797
    5. Yao Lu, Giulia Allegri, Jurriaan Huskens. Recruitment of Receptors and Ligands in a Weakly Multivalent System with Omnipresent Signatures of Superselective Binding. Small 2023, 19 (23) https://doi.org/10.1002/smll.202206596
    6. Shreya Pramanik, Jan Steinkühler, Rumiana Dimova, Joachim Spatz, Reinhard Lipowsky. Binding of His-tagged fluorophores to lipid bilayers of giant vesicles. Soft Matter 2022, 18 (34) , 6372-6383. https://doi.org/10.1039/D2SM00915C
    7. Sandra S. Flores, Pedro D. Clop, José L. Barra, Carlos E. Argaraña, María A. Perillo, Verónica Nolan, Julieta M. Sánchez. His-tag β-galactosidase supramolecular performance. Biophysical Chemistry 2022, 281 , 106739. https://doi.org/10.1016/j.bpc.2021.106739
    8. Kayla Sapp, Alexander J. Sodt. Observed steric crowding at modest coverage requires a particular membrane-binding scheme or a complementary mechanism. Biophysical Journal 2022, 121 (3) , 430-438. https://doi.org/10.1016/j.bpj.2021.12.036
    9. Maiara A. Iriarte-Alonso, Alexander M. Bittner, Salvatore Chiantia. Influenza A virus hemagglutinin prevents extensive membrane damage upon dehydration. BBA Advances 2022, 2 , 100048. https://doi.org/10.1016/j.bbadva.2022.100048
    10. Hèctor López-Laguna, Eric Voltà-Durán, Eloi Parladé, Antonio Villaverde, Esther Vázquez, Ugutz Unzueta. Insights on the emerging biotechnology of histidine-rich peptides. Biotechnology Advances 2022, 54 , 107817. https://doi.org/10.1016/j.biotechadv.2021.107817
    11. Bastiaan C. Buddingh', Antoni Llopis-Lorente, Loai K. E. A. Abdelmohsen, Jan C. M. van Hest. Dynamic spatial and structural organization in artificial cells regulates signal processing by protein scaffolding. Chemical Science 2020, 11 (47) , 12829-12834. https://doi.org/10.1039/D0SC03933K

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