Peptide-nucleotide microdroplets as a step towards a membrane-free protocell model.

Although phospholipid bilayers are ubiquitous in modern cells, their impermeability, lack of dynamic properties, and synthetic complexity are difficult to reconcile with plausible pathways of proto-metabolism, growth and division. Here, we present an alternative membrane-free model, which demonstrates that low-molecular-weight mononucleotides and simple cationic peptides spontaneously accumulate in water into microdroplets that are stable to changes in temperature and salt concentration, undergo pH-induced cycles of growth and decay, and promote α-helical peptide secondary structure. Moreover, the microdroplets selectively sequester porphyrins, inorganic nanoparticles and enzymes to generate supramolecular stacked arrays of light-harvesting molecules, nanoparticle-mediated oxidase activity, and enhanced rates of glucose phosphorylation, respectively. Taken together, our results suggest that peptide-nucleotide microdroplets can be considered as a new type of protocell model that could be used to develop novel bioreactors, primitive artificial cells and plausible pathways to prebiotic organization before the emergence of lipid-based compartmentalization on the early Earth.

[1]  P. Walde,et al.  Building artificial cells and protocell models: experimental approaches with lipid vesicles. , 2010, BioEssays : news and reviews in molecular, cellular and developmental biology.

[2]  G. Felsenfeld,et al.  Deoxyribonucleic acid-polylysine complexes. Structure and nucleotide specificity. , 1969, Biochemistry.

[3]  D. Deamer,et al.  Amphiphilic components of the murchison carbonaceous chondrite: Surface properties and membrane formation , 2005, Origins of life and evolution of the biosphere.

[4]  Kenichi Yoshikawa,et al.  Gene Expression within Cell‐Sized Lipid Vesicles , 2003, Chembiochem : a European journal of chemical biology.

[5]  P. Dubin,et al.  Entering and exiting the protein-polyelectrolyte coacervate phase via nonmonotonic salt dependence of critical conditions. , 2010, Biomacromolecules.

[6]  J. Oró,et al.  Possible prebiotic significance of polyamines in the condensation, protection, encapsulation, and biological properties of DNA , 2005, Origins of life and evolution of the biosphere.

[7]  Pier Luigi Luisi,et al.  The Emergence Of Life , 2006 .

[8]  Pier Luigi Luisi,et al.  OPARIN'S REACTIONS REVISITED : ENZYMATIC SYNTHESIS OF POLY(ADENYLIC ACID) IN MICELLES AND SELF-REPRODUCING VESICLES , 1994 .

[9]  A. Oparin [The origin of life]. , 1938, Nordisk medicin.

[10]  J. Pelta,et al.  DNA Aggregation Induced by Polyamines and Cobalthexamine (*) , 1996, The Journal of Biological Chemistry.

[11]  Cornelia Meinert,et al.  On the origin of primitive cells: from nutrient intake to elongation of encapsulated nucleotides. , 2010, Angewandte Chemie.

[12]  D. Bartel,et al.  Synthesizing life , 2001, Nature.

[13]  Charalambos Kaittanis,et al.  Oxidase-like activity of polymer-coated cerium oxide nanoparticles. , 2009, Angewandte Chemie.

[14]  G. Wächtershäuser,et al.  Evolution of the first metabolic cycles. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[15]  D. Bartel,et al.  Synthesizing life : Paths to unforeseeable science & technology , 2001 .

[16]  D. Deamer,et al.  Liposomes from ionic, single-chain amphiphiles. , 1978, Biochemistry.

[17]  S. Ichikawa,et al.  Enzymes inside lipid vesicles: preparation, reactivity and applications. , 2001, Biomolecular engineering.

[18]  A. Golestani,et al.  Interaction of hexokinase with the outer mitochondrial membrane and a hydrophobic matrix , 2001, Molecular and Cellular Biochemistry.

[19]  J. Waite,et al.  Promotion of osteoblast proliferation on complex coacervation-based hyaluronic acid - recombinant mussel adhesive protein coatings on titanium. , 2010, Biomaterials.

[20]  P. Luisi The Emergence of Life: Autopoiesis: the logic of cellular life , 2006 .

[21]  Yi‐Yeoun Kim,et al.  Controlled nanoparticle formation by enzymatic deshelling of biopolymer-stabilized nanosuspensions. , 2009, Small.

[22]  J. Ferris,et al.  Oligomerization of Ribonucleotides on Montmorillonite: Reaction of the 5′‐Phosphorimidazolide of Adenosine. , 1993 .

[23]  P. Luisi,et al.  Polymerase chain reaction in liposomes. , 1995, Chemistry & biology.

[24]  G. Fasman,et al.  Computed circular dichroism spectra for the evaluation of protein conformation. , 1969, Biochemistry.

[25]  J. Szostak,et al.  The origins of cellular life. , 2010, Cold Spring Harbor perspectives in biology.

[26]  Steen Rasmussen,et al.  Protocells : bridging nonliving and living matter , 2008 .

[27]  F. Weinbreck,et al.  Complex coacervation of proteins and anionic polysaccharides , 2004 .

[28]  Vincent Noireaux,et al.  A vesicle bioreactor as a step toward an artificial cell assembly. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[29]  J. Szostak,et al.  Template-directed synthesis of a genetic polymer in a model protocell , 2008, Nature.

[30]  B. Simoneit,et al.  Lipid Synthesis Under Hydrothermal Conditions by Fischer- Tropsch-Type Reactions , 1999, Origins of life and evolution of the biosphere.

[31]  Teruo Umemoto,et al.  Preparation, Reactivity, and Applications of N-Fluoropyridinium Salts , 2009 .

[32]  P. Luisi,et al.  Enzymatic RNA replication in self-reproducing vesicles: an approach to a minimal cell. , 1995, Biochemical and biophysical research communications.

[33]  I. A. Rose,et al.  Ascites tumor mitochondrial hexokinase II. Effect of binding on kinetic properties. , 1968, The Journal of biological chemistry.

[34]  S. Fox The evolutionary significance of phase-separated microsystems , 2005, Origins of life.