Mimicking Cellular Compartmentalization in a Hierarchical Protocell through Spontaneous Spatial Organization.

ACS Cent Sci. 2019 Aug 28; 5(8): 1360–1365. Published online 2019 Jul 3. doi: 10.1021/acscentsci.9b00345 PMCID: PMC6716124 PMID: 31482118 Mimicking Cellular Compartmentalization in a Hierarchical Protocell through Spontaneous Spatial Organization Alexander F. Mason,†# N. Amy Yewdall,†# Pascal L. W. Welzen,† Jingxin Shao,† Marleen van Stevendaal,† Jan C. M. van Hest,*† David S. Williams,*‡ and Loai K. E. A. Abdelmohsen*† Alexander F. Mason †Department of Biomedical Engineering & Department of Chemical Engineering and Chemistry, Institute for Complex Molecular Systems, Eindhoven University of Technology, P.O. Box 513, 5600 MB Eindhoven, The Netherlands Find articles by Alexander F. Mason N. Amy Yewdall †Department of Biomedical Engineering & Department of Chemical Engineering and Chemistry, Institute for Complex Molecular Systems, Eindhoven University of Technology, P.O. Box 513, 5600 MB Eindhoven, The Netherlands Find articles by N. Amy Yewdall Pascal L. W. Welzen †Department of Biomedical Engineering & Department of Chemical Engineering and Chemistry, Institute for Complex Molecular Systems, Eindhoven University of Technology, P.O. Box 513, 5600 MB Eindhoven, The Netherlands Find articles by Pascal L. W. Welzen Jingxin Shao †Department of Biomedical Engineering & Department of Chemical Engineering and Chemistry, Institute for Complex Molecular Systems, Eindhoven University of Technology, P.O. Box 513, 5600 MB Eindhoven, The Netherlands Find articles by Jingxin Shao Marleen van Stevendaal †Department of Biomedical Engineering & Department of Chemical Engineering and Chemistry, Institute for Complex Molecular Systems, Eindhoven University of Technology, P.O. Box 513, 5600 MB Eindhoven, The Netherlands Find articles by Marleen van Stevendaal Jan C. M. van Hest †Department of Biomedical Engineering & Department of Chemical Engineering and Chemistry, Institute for Complex Molecular Systems, Eindhoven University of Technology, P.O. Box 513, 5600 MB Eindhoven, The Netherlands Find articles by Jan C. M. van Hest David S. Williams ‡Department of Chemistry, College of Science, Swansea University, Singleton Campus, Swansea, Wales SA2 8PP, United Kingdom Find articles by David S. Williams Loai K. E. A. Abdelmohsen †Department of Biomedical Engineering & Department of Chemical Engineering and Chemistry, Institute for Complex Molecular Systems, Eindhoven University of Technology, P.O. Box 513, 5600 MB Eindhoven, The Netherlands Find articles by Loai K. E. A. Abdelmohsen Author information Article notes Copyright and License information Disclaimer †Department of Biomedical Engineering & Department of Chemical Engineering and Chemistry, Institute for Complex Molecular Systems, Eindhoven University of Technology, P.O. Box 513, 5600 MB Eindhoven, The Netherlands ‡Department of Chemistry, College of Science, Swansea University, Singleton Campus, Swansea, Wales SA2 8PP, United Kingdom Corresponding author. *(J.C.M.v.H.) E-mail: [email protected]. *(D.S.W.) E-mail: [email protected]. *(L.K.E.A.A.) E-mail: [email protected]. Received 2019 Apr 4 Copyright © 2019 American Chemical Society This is an open access article published under a Creative Commons Non-Commercial No Derivative Works (CC-BY-NC-ND) Attribution License, which permits copying and redistribution of the article, and creation of adaptations, all for non-commercial purposes. Associated DataSupplementary Materials oc9b00345_si_001.pdf oc9b00345_si_001.pdf (2.3M) GUID: 4A271401-705C-4DE4-B66C-1F8CC87A458A Abstract Open in a separate window A systemic feature of eukaryotic cells is the spatial organization of functional components through compartmentalization. Developing protocells with compartmentalized synthetic organelles is, therefore, a critical milestone toward emulating one of the core characteristics of cellular life. Here we demonstrate the bottom-up, multistep, noncovalent, assembly of rudimentary subcompartmentalized protocells through the spontaneous encapsulation of semipermeable, polymersome proto-organelles inside cell-sized coacervates. The coacervate microdroplets are membranized using tailor-made terpolymers, to complete the hierarchical self-assembly of protocells, a system that mimics both the condensed cytosol and the structure of a cell membrane. In this way, the spatial organization of enzymes can be finely tuned, leading to an enhancement of functionality. Moreover, incompatible components can be sequestered in the same microenvironments without detrimental effect. The robust stability of the subcompartmentalized coacervate protocells in biocompatible milieu, such as in PBS or cell culture media, makes it a versatile platform to be extended toward studies in vitro, and perhaps, in vivo. Short abstract Herein, hierarchical protocells are engineered by multistep, noncovalent assembly generating subcompartmentalized, membranized, coacervates—a functional model of eukaryotic spatial organization. Introduction Compartmentalization is key for the emergence of eukaryotic life, facilitating stepwise enhancements in complexity toward increasingly functional forms of hierarchically structured matter.1,2 With growing interest in the development of synthetic cell-like architectures (protocells), various forms of micro- and nanostructures that mimic essential cellular properties and processes have been presented.3−5 To date, protocell research has largely focused on the development of discrete chemical platforms such as membrane-free protocells (coacervates,6−11 hydrogel particles,12,13 or aqueous two-phase systems14,15) and membrane-bound protocells (liposomes,16 proteinosomes,8 colloidosomes,5,17 polymeric nanoparticles,18 or polymersomes19,20). These first-generation protocells have been implemented to advance fundamental understanding into the physicochemical hallmarks of living systems.21 With an emphasis on the engineering of hybrid systems, next-generation protocells seek to take this a step further, increasing structural and functional complexity by implementing multicompartmentalization.22−28 Exemplifying this, bottom-up engineering of cell-mimetic liposomes (loaded with enzymes)29 have been subcompartmentalized within polymeric particles, displaying temperature-dependent localization.30 Top-down approaches have also been adopted, bridging the divide between natural and synthetic systems by incorporating biological organelles within protocells31 and, conversely, amalgamating synthetic organelles with cells, imparting new functionality.32−36 The hierarchical assembly of subcompartmentalized systems is a key milestone in the development of protocells—a process that showcases the multistep, noncovalent assembly of complex materials that resemble lifelike systems.37 Here, we present a multicompartmentalized, coacervate-based protocell capable of spontaneously assimilating new functional elements, in a proce (...truncated)