Controlling Protein Enrichment in Lipid Sponge Phase Droplets using SNAP–Tag Bioconjugation

Moinpour, Mahta; Fracassi, Alessandro; Brea, Roberto J.; Salvador‐Castell, Marta; Pandey, Sudip; Edwards, Madison M.; Seifert, Soenke; Joseph, Simpson; Sinha, Sunil K.; Devaraj, Neal K.

doi:10.1002/cbic.202100624

Title: Controlling Protein Enrichment in Lipid Sponge Phase Droplets using SNAP–Tag Bioconjugation

Journal Article · 19 January 2022 · ChemBioChem: a European journal of chemical biology

DOI: https://doi.org/10.1002/cbic.202100624 · OSTI ID:1879855

Moinpour, Mahta ^[1];

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Univ. of California, San Diego, La Jolla, CA (United States)
Universidade da Coruña (Spain)
Argonne National Lab. (ANL), Lemont, IL (United States)

All cells use organized lipid compartments to facilitate specific biological functions. Membrane-bound organelles create defined spatial environments that favor unique chemical reactions while isolating incompatible biological processes. Despite the fundamental role of cellular organelles, there is a scarcity of methods for preparing functional artificial lipid-based compartments. In this work, we demonstrate a robust bioconjugation system for sequestering proteins into zwitterionic lipid sponge phase droplets. Incorporation of benzylguanine (BG)-modified phospholipids that form stable covalent linkages with an O⁶-methylguanine DNA methyltransferase (SNAP-tag) fusion protein enables programmable control of protein capture. We show that this methodology can be used to anchor hydrophilic proteins at the lipid-aqueous interface, concentrating them within an accessible but protected chemical environment. SNAP-tag technology enables the integration of proteins that regulate complex biological functions in lipid sponge phase droplets, and should facilitate the development of advanced lipid-based artificial organelles.

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Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES); US Army Research Office (ARO); Xunta de Galicia; University of California

Grant/Contract Number:: AC02-06CH11357

OSTI ID:: 1879855

Journal Information:: ChemBioChem: a European journal of chemical biology, Journal Name: ChemBioChem: a European journal of chemical biology Journal Issue: 5 Vol. 23; ISSN 1439-4227

Publisher:: ChemPubSoc EuropeCopyright Statement

Country of Publication:: United States

Language:: English

References (47)

Oberflächenfunktionalisierung durch spannungsvermittelte Alkin-Azid-Klickreaktionen Manova, Radostina; van Beek, Teris A.; Zuilhof, Han Angewandte Chemie, Vol. 123, Issue 24 https://doi.org/10.1002/ange.201100835	journal	May 2011
Phase Behavior of a Designed Cyclopropyl Analogue of Monoolein: Implications for Low-Temperature Membrane Protein Crystallization Salvati Manni, Livia; Zabara, Alexandru; Osornio, Yazmin M. Angewandte Chemie, Vol. 127, Issue 3 https://doi.org/10.1002/ange.201409791	journal	November 2014
Nanostructure Engineering by Simple Tuning of Lipid Combinations Saw, Phei Er; Xu, Xiaoding; Zhang, Meng Angewandte Chemie, Vol. 132, Issue 15 https://doi.org/10.1002/ange.201916574	journal	February 2020
Surface Functionalization by Strain-Promoted Alkyne-Azide Click Reactions Manova, Radostina; van Beek, Teris A.; Zuilhof, Han Angewandte Chemie International Edition, Vol. 50, Issue 24 https://doi.org/10.1002/anie.201100835	journal	May 2011
Phase Behavior of a Designed Cyclopropyl Analogue of Monoolein: Implications for Low-Temperature Membrane Protein Crystallization Salvati Manni, Livia; Zabara, Alexandru; Osornio, Yazmin M. Angewandte Chemie International Edition, Vol. 54, Issue 3 https://doi.org/10.1002/anie.201409791	journal	November 2014
Nanostructure Engineering by Simple Tuning of Lipid Combinations Saw, Phei Er; Xu, Xiaoding; Zhang, Meng Angewandte Chemie International Edition, Vol. 59, Issue 15 https://doi.org/10.1002/anie.201916574	journal	February 2020
The GPI-anchor and protein sorting Chatterjee, S.; Mayor, S. Cellular and Molecular Life Sciences, Vol. 58, Issue 14 https://doi.org/10.1007/PL00000831	journal	December 2001
Membrane fusion and molecular segregation in phospholipid vesicles Papahadjopoulos, D.; Poste, G.; Schaeffer, B. E. Biochimica et Biophysica Acta (BBA) - Biomembranes, Vol. 352, Issue 1 https://doi.org/10.1016/0005-2736(74)90175-8	journal	May 1974
X-ray diffraction studies of lecithin bilayers Torbet, J.; Wilkins, M. H. F. Journal of Theoretical Biology, Vol. 62, Issue 2 https://doi.org/10.1016/0022-5193(76)90129-6	journal	October 1976
Principles and applications of grazing incidence X-ray and neutron scattering from ordered molecular monolayers at the air-water interface Als-Nielsen, Jens; Jacquemain, Didier; Kjaer, Kristian Physics Reports, Vol. 246, Issue 5 https://doi.org/10.1016/0370-1573(94)90046-9	journal	October 1994
Quantitation of lipid phases in phospholipid vesicles by the generalized polarization of Laurdan fluorescence Parasassi, T.; De Stasio, G.; Ravagnan, G. Biophysical Journal, Vol. 60, Issue 1 https://doi.org/10.1016/S0006-3495(91)82041-0	journal	July 1991
Lipid-dependent protein sorting at the trans-Golgi network Surma, Michal A.; Klose, Christian; Simons, Kai Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids, Vol. 1821, Issue 8 https://doi.org/10.1016/j.bbalip.2011.12.008	journal	August 2012
Cholesterol Partition and Condensing Effect in Phase-Separated Ternary Mixture Lipid Multilayers Ma, Yicong; Ghosh, Sajal K.; DiLena, David A. Biophysical Journal, Vol. 110, Issue 6 https://doi.org/10.1016/j.bpj.2016.02.022	journal	March 2016
Membrane mimetic surface functionalization of nanoparticles: Methods and applications Weingart, Jacob; Vabbilisetty, Pratima; Sun, Xue-Long Advances in Colloid and Interface Science, Vol. 197-198 https://doi.org/10.1016/j.cis.2013.04.003	journal	September 2013
Phospholipid-based nonlamellar mesophases for delivery systems: Bridging the gap between empirical and rational design Martiel, Isabelle; Sagalowicz, Laurent; Mezzenga, Raffaele Advances in Colloid and Interface Science, Vol. 209 https://doi.org/10.1016/j.cis.2014.03.005	journal	July 2014
How to obtain labeled proteins and what to do with them Hinner, Marlon J.; Johnsson, Kai Current Opinion in Biotechnology, Vol. 21, Issue 6 https://doi.org/10.1016/j.copbio.2010.09.011	journal	December 2010
Interface tuning and stabilization of monoglyceride mesophase dispersions: Food emulsifiers and mixtures efficiency Serieye, Sébastien; Méducin, Fabienne; Milošević, Irena Journal of Colloid and Interface Science, Vol. 496 https://doi.org/10.1016/j.jcis.2017.01.059	journal	June 2017
Design of Light-Triggered Lyotropic Liquid Crystal Mesophases and Their Application as Molecular Switches in “On Demand” Release Aleandri, Simone; Speziale, Chiara; Mezzenga, Raffaele Langmuir, Vol. 31, Issue 25 https://doi.org/10.1021/acs.langmuir.5b01945	journal	June 2015
Biotinylated Cubosomes: A Versatile Tool for Active Targeting and Codelivery of Paclitaxel and a Fluorescein-Based Lipid Dye Aleandri, Simone; Bandera, Davide; Mezzenga, Raffaele Langmuir, Vol. 31, Issue 46 https://doi.org/10.1021/acs.langmuir.5b03469	journal	November 2015
KAT Ligation for Rapid and Facile Covalent Attachment of Biomolecules to Surfaces Fracassi, Alessandro; Ray, Ankita; Nakatsuka, Nako ACS Applied Materials & Interfaces, Vol. 13, Issue 24 https://doi.org/10.1021/acsami.1c05652	journal	June 2021
Biomedical Applications of Tetrazine Cycloadditions Devaraj, Neal K.; Weissleder, Ralph Accounts of Chemical Research, Vol. 44, Issue 9 https://doi.org/10.1021/ar200037t	journal	May 2011
Coacervate Formation from Natural Glycolipid: One Acetyl Group on the Headgroup Triggers Coacervate-to-Vesicle Transition Imura, T.; Yanagishita, H.; Kitamoto, D. Journal of the American Chemical Society, Vol. 126, Issue 35 https://doi.org/10.1021/ja0400281	journal	September 2004
Membrane Mimetic Chemistry in Artificial Cells Vance, Jacob A.; Devaraj, Neal K. Journal of the American Chemical Society, Vol. 143, Issue 22 https://doi.org/10.1021/jacs.1c03436	journal	May 2021
SNAP-Tag-Reactive Lipid Anchors Enable Targeted and Spatiotemporally Controlled Localization of Proteins to Phospholipid Membranes Rudd, Andrew K.; Valls Cuevas, Joan M.; Devaraj, Neal K. Journal of the American Chemical Society, Vol. 137, Issue 15 https://doi.org/10.1021/jacs.5b00040	journal	April 2015
A Sponge Morphology in an Elementary Coacervate Menger, F. M.; Peresypkin, A. V.; Caran, K. L. Langmuir, Vol. 16, Issue 24 https://doi.org/10.1021/la0010626	journal	November 2000
Scaling of Structural and Rheological Response of L ₃ Sponge Phases in the “Sweetened” Cetylpyridinium/Hexanol/Dextrose/Brine System Porcar, L.; Hamilton, W. A.; Butler, P. D. Langmuir, Vol. 19, Issue 26 https://doi.org/10.1021/la034661w	journal	December 2003
Zwitterionic Geminis. Coacervate Formation from a Single Organic Compound Peresypkin, Andrey V.; Menger, Fredric M. Organic Letters, Vol. 1, Issue 9 https://doi.org/10.1021/ol990205g	journal	November 1999
Lipid Metabolism and Regulation of Membrane Trafficking: Lipid Metabolism and Regulation of Membrane Trafficking Huijbregts, Richard P. H.; Topalof, Lora; Bankaitis, Vytas A. Traffic, Vol. 1, Issue 3 https://doi.org/10.1034/j.1600-0854.2000.010301.x	journal	March 2000
Cell-free translation reconstituted with purified components Shimizu, Yoshihiro; Inoue, Akio; Tomari, Yukihide Nature Biotechnology, Vol. 19, Issue 8 https://doi.org/10.1038/90802	journal	August 2001
Photosynthetic artificial organelles sustain and control ATP-dependent reactions in a protocellular system Lee, Keel Yong; Park, Sung-Jin; Lee, Keon Ah Nature Biotechnology, Vol. 36, Issue 6 https://doi.org/10.1038/nbt.4140	journal	May 2018
A general method for the covalent labeling of fusion proteins with small molecules in vivo Keppler, Antje; Gendreizig, Susanne; Gronemeyer, Thomas Nature Biotechnology, Vol. 21, Issue 1 https://doi.org/10.1038/nbt765	journal	December 2002
Size-dependent protein segregation at membrane interfaces Schmid, Eva M.; Bakalar, Matthew H.; Choudhuri, Kaushik Nature Physics, Vol. 12, Issue 7 https://doi.org/10.1038/nphys3678	journal	March 2016
Biology under construction: in vitro reconstitution of cellular function Liu, Allen P.; Fletcher, Daniel A. Nature Reviews Molecular Cell Biology, Vol. 10, Issue 9 https://doi.org/10.1038/nrm2746	journal	August 2009
Design of ultra-swollen lipidic mesophases for the crystallization of membrane proteins with large extracellular domains Zabara, Alexandru; Chong, Josephine Tse Yin; Martiel, Isabelle Nature Communications, Vol. 9, Issue 1 https://doi.org/10.1038/s41467-018-02996-5	journal	February 2018
Synthesis of lipid membranes for artificial cells Podolsky, Kira A.; Devaraj, Neal K. Nature Reviews Chemistry, Vol. 5, Issue 10 https://doi.org/10.1038/s41570-021-00303-3	journal	July 2021
SAXS investigation of a cubic to a sponge (L ₃ ) phase transition in self-assembled lipid nanocarriers Angelov, Borislav; Angelova, Angelina; Mutafchieva, Rada Phys. Chem. Chem. Phys., Vol. 13, Issue 8 https://doi.org/10.1039/C0CP01029D	journal	January 2011
Lipid-based mesophases as matrices for nanoscale reactions Salvati Manni, Livia; Fong, Wye-Khay; Mezzenga, Raffaele Nanoscale Horizons, Vol. 5, Issue 6 https://doi.org/10.1039/D0NH00079E	journal	January 2020
LDL-mimetic lipid nanoparticles prepared by surface KAT ligation for in vivo MRI of atherosclerosis Fracassi, Alessandro; Cao, Jianbo; Yoshizawa-Sugata, Naoko Chemical Science, Vol. 11, Issue 44 https://doi.org/10.1039/D0SC04106H	journal	January 2020
The physics of lipidic mesophase delivery systems Aleandri, Simone; Mezzenga, Raffaele Physics Today, Vol. 73, Issue 7 https://doi.org/10.1063/PT.3.4522	journal	July 2020
Lipid sponge droplets as programmable synthetic organelles Bhattacharya, Ahanjit; Niederholtmeyer, Henrike; Podolsky, Kira A. Proceedings of the National Academy of Sciences, Vol. 117, Issue 31 https://doi.org/10.1073/pnas.2004408117	journal	July 2020
Lipidic cubic phases: A novel concept for the crystallization of membrane proteins Landau, E. M.; Rosenbusch, J. P. Proceedings of the National Academy of Sciences, Vol. 93, Issue 25 https://doi.org/10.1073/pnas.93.25.14532	journal	December 1996
Cubic membranes: a legend beyond the Flatland* of cell membrane organization Almsherqi, Zakaria A.; Kohlwein, Sepp D.; Deng, Yuru The Journal of Cell Biology, Vol. 173, Issue 6 https://doi.org/10.1083/jcb.200603055	journal	June 2006
Cubic membranes: a structure-based design for DNA uptake Almsherqi, Zakaria; Hyde, Stephen; Ramachandran, Malarmathy Journal of The Royal Society Interface, Vol. 5, Issue 26 https://doi.org/10.1098/rsif.2007.1351	journal	February 2008
Swelling behavior and local topology of an L 3 (sponge) phase Maldonado, A.; Urbach, W.; Ober, R. Physical Review E, Vol. 54, Issue 2 https://doi.org/10.1103/PhysRevE.54.1774	journal	August 1996
Synchrotron x-ray-scattering studies on the sodium dodecyl sulfate–water–pentanol–dodecane L 3 sponge phase Lei, Ning; Safinya, C. R.; Roux, D. Physical Review E, Vol. 56, Issue 1 https://doi.org/10.1103/PhysRevE.56.608	journal	July 1997
Membrane topology of Escherichia coli diacylglycerol kinase. Smith, R. L.; O'Toole, J. F.; Maguire, M. E. Journal of Bacteriology, Vol. 176, Issue 17 https://doi.org/10.1128/jb.176.17.5459-5465.1994	journal	September 1994
Mechanisms Shaping the Membranes of Cellular Organelles Shibata, Yoko; Hu, Junjie; Kozlov, Michael M. Annual Review of Cell and Developmental Biology, Vol. 25, Issue 1 https://doi.org/10.1146/annurev.cellbio.042308.113324	journal	November 2009

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59 BASIC BIOLOGICAL SCIENCES
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Title: Controlling Protein Enrichment in Lipid Sponge Phase Droplets using SNAP–Tag Bioconjugation

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