Lipid sponge droplets as programmable synthetic organelles

Bhattacharya, Ahanjit; Niederholtmeyer, Henrike; Podolsky, Kira A.; Bhattacharya, Rupak; Song, Jing-Jin; Brea, Roberto J.; Tsai, Chu-Hsien; Sinha, Sunil K.; Devaraj, Neal K.

doi:10.1073/pnas.2004408117

Lipid sponge droplets as programmable synthetic organelles

Journal Article · Tue Jul 21 00:00:00 EDT 2020 · Proceedings of the National Academy of Sciences of the United States of America

DOI:https://doi.org/10.1073/pnas.2004408117· OSTI ID:1640278

; ; ; Bhattacharya, Rupak; Song, Jing-Jin; ; Tsai, Chu-Hsien; Sinha, Sunil K.;

Living cells segregate molecules and reactions in various subcellular compartments known as organelles. Spatial organization is likely essential for expanding the biochemical functions of synthetic reaction systems, including artificial cells. Many studies have attempted to mimic organelle functions using lamellar membrane-bound vesicles. However, vesicles typically suffer from highly limited transport across the membranes and an inability to mimic the dense membrane networks typically found in organelles such as the endoplasmic reticulum. Here, we describe programmable synthetic organelles based on highly stable nonlamellar sponge phase droplets that spontaneously assemble from a single-chain galactolipid and nonionic detergents. Due to their nanoporous structure, lipid sponge droplets readily exchange materials with the surrounding environment. In addition, the sponge phase contains a dense network of lipid bilayers and nanometric aqueous channels, which allows different classes of molecules to partition based on their size, polarity, and specific binding motifs. The sequestration of biologically relevant macromolecules can be programmed by the addition of suitably functionalized amphiphiles to the droplets. We demonstrate that droplets can harbor functional soluble and transmembrane proteins, allowing for the colocalization and concentration of enzymes and substrates to enhance reaction rates. Droplets protect bound proteins from proteases, and these interactions can be engineered to be reversible and optically controlled. Our results show that lipid sponge droplets permit the facile integration of membrane-rich environments and self-assembling spatial organization with biochemical reaction systems.

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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). Materials Sciences & Engineering Division

Contributing Organization:: Advanced Photon Source (APS), Argonne National Laboratory (ANL), Argonne, IL (US)

Grant/Contract Number:: SC0018086

OSTI ID:: 1640278

Alternate ID(s):: OSTI ID: 1671134

Journal Information:: Proceedings of the National Academy of Sciences of the United States of America, Journal Name: Proceedings of the National Academy of Sciences of the United States of America Journal Issue: 31 Vol. 117; ISSN 0027-8424

Publisher:: National Academy of SciencesCopyright Statement

Country of Publication:: United States

Language:: English

References (52)

Cascade Reactions in Multicompartmentalized Polymersomes Peters, Ruud J. R. W.; Marguet, Maïté; Marais, Sébastien Angewandte Chemie International Edition, Vol. 53, Issue 1 https://doi.org/10.1002/anie.201308141	journal	November 2013
Sequestration of Proteins by Fatty Acid Coacervates for Their Encapsulation within Vesicles Garenne, David; Beven, Laure; Navailles, Laurence Angewandte Chemie International Edition, Vol. 55, Issue 43 https://doi.org/10.1002/anie.201607117	journal	September 2016
Microfluidic Formation of Monodisperse Coacervate Organelles in Liposomes Deng, Nan-Nan; Huck, Wilhelm T. S. Angewandte Chemie International Edition, Vol. 56, Issue 33 https://doi.org/10.1002/anie.201703145	journal	July 2017
Artificial Gel-Based Organelles for Spatial Organization of Cell-Free Gene Expression Reactions Aufinger, Lukas; Simmel, Friedrich C. Angewandte Chemie International Edition, Vol. 57, Issue 52 https://doi.org/10.1002/anie.201809374	journal	November 2018
Fatty Acid/Phospholipid Blended Membranes: A Potential Intermediate State in Protocellular Evolution Jin, Lin; Kamat, Neha P.; Jena, Siddhartha Small, Vol. 14, Issue 15 https://doi.org/10.1002/smll.201704077	journal	February 2018
The endoplasmic reticulum: structure, function and response to cellular signaling Schwarz, Dianne S.; Blower, Michael D. Cellular and Molecular Life Sciences, Vol. 73, Issue 1 https://doi.org/10.1007/s00018-015-2052-6	journal	October 2015
Generic and reversible opto-trapping of biomolecules Beyer, Hannes M.; Thomas, Oliver S.; Riegel, Nils Acta Biomaterialia, Vol. 79 https://doi.org/10.1016/j.actbio.2018.08.032	journal	October 2018
Spatiotemporal Control of Intracellular Phase Transitions Using Light-Activated optoDroplets Shin, Yongdae; Berry, Joel; Pannucci, Nicole Cell, Vol. 168, Issue 1-2 https://doi.org/10.1016/j.cell.2016.11.054	journal	January 2017
Structural Analysis of Lac Repressor Bound to Allosteric Effectors Daber, Robert; Stayrook, Steven; Rosenberg, Allison Journal of Molecular Biology, Vol. 370, Issue 4 https://doi.org/10.1016/j.jmb.2007.04.028	journal	July 2007
Spatiotemporal organization of microbial cells by protein concentration gradients Kiekebusch, Daniela; Thanbichler, Martin Trends in Microbiology, Vol. 22, Issue 2 https://doi.org/10.1016/j.tim.2013.11.005	journal	February 2014
Single-Chain β- d -Glycopyranosylamides of Unsaturated Fatty Acids: Self-Assembly Properties and Applications to Artificial Cell Development Bhattacharya, Ahanjit; Brea, Roberto J.; Song, Jing-Jin The Journal of Physical Chemistry B, Vol. 123, Issue 17 https://doi.org/10.1021/acs.jpcb.9b01055	journal	April 2019
Characterization of a Biomimetic Mesophase Composed of Nonionic Surfactants and an Aqueous Solvent Adrien, V.; Rayan, G.; Reffay, M. Langmuir, Vol. 32, Issue 40 https://doi.org/10.1021/acs.langmuir.6b02744	journal	September 2016
Coassembly of Photosystem II and ATPase as Artificial Chloroplast for Light-Driven ATP Synthesis Feng, Xiyun; Jia, Yi; Cai, Peng ACS Nano, Vol. 10, Issue 1 https://doi.org/10.1021/acsnano.5b05579	journal	December 2015
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
Bottom-up Construction of a Primordial Carboxysome Mimic Frey, Raphael; Mantri, Shiksha; Rocca, Marco Journal of the American Chemical Society, Vol. 138, Issue 32 https://doi.org/10.1021/jacs.6b04744	journal	August 2016
Harnessing Yeast Peroxisomes for Biosynthesis of Fatty-Acid-Derived Biofuels and Chemicals with Relieved Side-Pathway Competition Zhou, Yongjin J.; Buijs, Nicolaas A.; Zhu, Zhiwei Journal of the American Chemical Society, Vol. 138, Issue 47 https://doi.org/10.1021/jacs.6b07394	journal	October 2016
Microfluidic Assembly of Monodisperse Vesosomes as Artificial Cell Models Deng, Nan-Nan; Yelleswarapu, Maaruthy; Zheng, Lifei Journal of the American Chemical Society, Vol. 139, Issue 2 https://doi.org/10.1021/jacs.6b10977	journal	December 2016
Hierarchical Self-Assembly of a Copolymer-Stabilized Coacervate Protocell Mason, Alexander F.; Buddingh’, Bastiaan C.; Williams, David S. Journal of the American Chemical Society, Vol. 139, Issue 48 https://doi.org/10.1021/jacs.7b10846	journal	November 2017
Freeze fracture electron microscopy of dilute lamellar and anomalous isotropic (L3) phases Strey, Reinhard; Jahn, W.; Porte, G. Langmuir, Vol. 6, Issue 11 https://doi.org/10.1021/la00101a003	journal	November 1990
Anatomy of a Coacervate Menger, Fredric M.; Sykes, Bridget M. Langmuir, Vol. 14, Issue 15 https://doi.org/10.1021/la980208m	journal	July 1998
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
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
Compartmentalization of metabolic pathways in yeast mitochondria improves the production of branched-chain alcohols Avalos, José L.; Fink, Gerald R.; Stephanopoulos, Gregory Nature Biotechnology, Vol. 31, Issue 4 https://doi.org/10.1038/nbt.2509	journal	February 2013
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
Peptide–nucleotide microdroplets as a step towards a membrane-free protocell model Koga, Shogo; Williams, David S.; Perriman, Adam W. Nature Chemistry, Vol. 3, Issue 9 https://doi.org/10.1038/nchem.1110	journal	August 2011
Fatty acid membrane assembly on coacervate microdroplets as a step towards a hybrid protocell model Dora Tang, T-Y.; Rohaida Che Hak, C.; Thompson, Alexander J. Nature Chemistry, Vol. 6, Issue 6 https://doi.org/10.1038/nchem.1921	journal	April 2014
Phosphorylation-mediated RNA/peptide complex coacervation as a model for intracellular liquid organelles Aumiller, William M.; Keating, Christine D. Nature Chemistry, Vol. 8, Issue 2 https://doi.org/10.1038/nchem.2414	journal	December 2015
Interfacial assembly of protein–polymer nano-conjugates into stimulus-responsive biomimetic protocells Huang, Xin; Li, Mei; Green, David C. Nature Communications, Vol. 4, Issue 1 https://doi.org/10.1038/ncomms3239	journal	July 2013
Sequential bottom-up assembly of mechanically stabilized synthetic cells by microfluidics Weiss, Marian; Frohnmayer, Johannes Patrick; Benk, Lucia Theresa Nature Materials, Vol. 17, Issue 1 https://doi.org/10.1038/nmat5005	journal	October 2017
Light-triggered enzymatic reactions in nested vesicle reactors Hindley, James W.; Elani, Yuval; McGilvery, Catriona M. Nature Communications, Vol. 9, Issue 1 https://doi.org/10.1038/s41467-018-03491-7	journal	March 2018
Communication and quorum sensing in non-living mimics of eukaryotic cells Niederholtmeyer, Henrike; Chaggan, Cynthia; Devaraj, Neal K. Nature Communications, Vol. 9, Issue 1 https://doi.org/10.1038/s41467-018-07473-7	journal	November 2018
Template-directed RNA polymerization and enhanced ribozyme catalysis inside membraneless compartments formed by coacervates Poudyal, Raghav R.; Guth-Metzler, Rebecca M.; Veenis, Andrew J. Nature Communications, Vol. 10, Issue 1 https://doi.org/10.1038/s41467-019-08353-4	journal	January 2019
Cryo-EM structure of the ClpXP protein degradation machinery Gatsogiannis, Christos; Balogh, Dora; Merino, Felipe Nature Structural & Molecular Biology, Vol. 26, Issue 10 https://doi.org/10.1038/s41594-019-0304-0	journal	October 2019
Optimization of ClpXP activity and protein synthesis in an E. coli extract-based cell-free expression system Shi, Xinying; Wu, Ti; M. Cole, Christian Scientific Reports, Vol. 8, Issue 1 https://doi.org/10.1038/s41598-018-21739-6	journal	February 2018
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
Interfacial properties of lipid sponge-like nanoparticles and the role of stabilizer on particle structure and surface interactions Valldeperas, Maria; Dabkowska, Aleksandra P.; Pálsson, Gunnar K. Soft Matter, Vol. 15, Issue 10 https://doi.org/10.1039/C8SM02634C	journal	January 2019
Dilute lamellar and L ₃ phases in the binary water–C ₁₂ E ₅ system Strey, Reinhard; Schomäcker, Reinhard; Roux, Didier J. Chem. Soc., Faraday Trans., Vol. 86, Issue 12 https://doi.org/10.1039/FT9908602253	journal	January 1990
DNA-functionalized hydrogels for confined membrane-free in vitro transcription/translation Thiele, J.; Ma, Y.; Foschepoth, D. Lab on a Chip, Vol. 14, Issue 15 https://doi.org/10.1039/c3lc51427g	journal	January 2014
Nanostructured bicontinuous cubic lipid self-assembly materials as matrices for protein encapsulation Conn, Charlotte E.; Drummond, Calum J. Soft Matter, Vol. 9, Issue 13 https://doi.org/10.1039/c3sm27743g	journal	January 2013
Synthetic organelles Simmel, Friedrich C. Emerging Topics in Life Sciences, Vol. 3, Issue 5 https://doi.org/10.1042/ETLS20190056	journal	August 2019
Enhanced transcription rates in membrane-free protocells formed by coacervation of cell lysate Sokolova, E.; Spruijt, E.; Hansen, M. M. K. Proceedings of the National Academy of Sciences, Vol. 110, Issue 29 https://doi.org/10.1073/pnas.1222321110	journal	July 2013
Lamellar phases and disordered phases of fluid bilayer membranes Porte, G. Journal of Physics: Condensed Matter, Vol. 4, Issue 45 https://doi.org/10.1088/0953-8984/4/45/002	journal	November 1992
Strategies for carbohydrate recognition by the mannose 6-phosphate receptors Dahms, N. M.; Olson, L. J.; Kim, J. -J. P. Glycobiology, Vol. 18, Issue 9 https://doi.org/10.1093/glycob/cwn061	journal	June 2008
Structural information from multilamellar liposomes at full hydration: Full q -range fitting with high quality x-ray data Pabst, Georg; Rappolt, Michael; Amenitsch, Heinz Physical Review E, Vol. 62, Issue 3 https://doi.org/10.1103/PhysRevE.62.4000	journal	September 2000
The Pseudomonas aeruginosa Lectins PA-IL and PA-IIL Are Controlled by Quorum Sensing and by RpoS Winzer, Klaus; Falconer, Colin; Garber, Nachman C. Journal of Bacteriology, Vol. 182, Issue 22 https://doi.org/10.1128/JB.182.22.6401-6411.2000	journal	November 2000
Liquid-Liquid Phase Separation in Biology Hyman, Anthony A.; Weber, Christoph A.; Jülicher, Frank Annual Review of Cell and Developmental Biology, Vol. 30, Issue 1 https://doi.org/10.1146/annurev-cellbio-100913-013325	journal	October 2014
Interactive Evolution of a Bicontinuous Structure Stenger, Florian; Voigt, Axel Leonardo, Vol. 53, Issue 3 https://doi.org/10.1162/leon_a_01614	journal	May 2020
Cell Biology by the Numbers Milo, Ron; Phillips, Rob https://doi.org/10.1201/9780429258770	book	December 2015
Galectins at a glance Johannes, Ludger; Jacob, Ralf; Leffler, Hakon Journal of Cell Science, Vol. 131, Issue 9 https://doi.org/10.1242/jcs.208884	journal	May 2018
Biomolecular Chemistry in Liquid Phase Separated Compartments Nakashima, Karina K.; Vibhute, Mahesh A.; Spruijt, Evan Frontiers in Molecular Biosciences, Vol. 6 https://doi.org/10.3389/fmolb.2019.00021	journal	April 2019
The nucleolar detention pathway: A cellular strategy for regulating molecular networks Audas, Timothy E.; Jacob, Mathieu D.; Lee, Stephen Cell Cycle, Vol. 11, Issue 11 https://doi.org/10.4161/cc.20140	journal	June 2012
Global, quantitative and dynamic mapping of protein subcellular localization Itzhak, Daniel N.; Tyanova, Stefka; Cox, Jürgen eLife, Vol. 5 https://doi.org/10.7554/eLife.16950	journal	June 2016

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