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A synthetic biology approach to the construction of membrane proteins in semi-synthetic minimal cells

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Kuruma, Yutetsu, Stano, Pasquale, Ueda, Takuya, Luisi, Pier Luigi·2009·Biochimica et Biophysica Acta (BBA) - Biomembranes

Abstract

Synthetic biology is an emerging field that aims at constructing artificial biological systems by combining engineering and molecular biology approaches. One of the most ambitious research line concerns the so-called semi-synthetic minimal cells, which are liposome-based system capable of synthesizing the lipids within the liposome surface. This goal can be reached by reconstituting membrane proteins within liposomes and allow them to synthesize lipids. This approach, that can be defined as biochemical, was already reported by us (Schmidli et al. J. Am. Chem. Soc. 113, 8127-8130, 1991). In more advanced models, however, a full reconstruction of the biochemical pathway requires (1) the synthesis of functional membrane enzymes inside liposomes, and (2) the local synthesis of lipids as catalyzed by the in situ synthesized enzymes. Here we show the synthesis and the activity--inside liposomes--of two membrane proteins involved in phospholipids biosynthesis pathway. The proteins, sn-glycerol-3-phosphate acyltransferase (GPAT) and lysophosphatidic acid acyltransferase (LPAAT), have been synthesized by using a totally reconstructed cell-free system (PURE system) encapsulated in liposomes. The activities of internally synthesized GPAT and LPAAT were confirmed by detecting the produced lysophosphatidic acid and phosphatidic acid, respectively. Through this procedure, we have implemented the first phase of a design aimed at synthesizing phospholipid membrane from liposome within from within - which corresponds to the autopoietic growth mechanism.

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Research works printed but not linked: 14

  1. Luisi, 2006, Approaches to semi-synthetic minimal cells: a review, Naturwissenschaften, 93, 1, 10.1007/s00114-005-0056-z · DOI · Paper page
  2. Forster, 2006, Towards synthesis of a minimal cell, Mol. Systems Biol., 2, 45, 10.1038/msb4100090 · DOI · Paper page
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  4. Schmidli, 1991, Liposome-mediated enzymatic synthesis of phosphatidylcholine as an approach to self-replicating liposome, J. Am. Chem. Soc., 113, 8127, 10.1021/ja00021a043 · DOI
  5. Shimizu, 2001, Cell-free translation reconstituted with purified components, Nature Biotechnol., 19, 751, 10.1038/90802 · DOI · Paper page
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  10. Murtas, 2007, Protein synthesis in liposomes with a minimal set of enzymes, Biochem. Biophys. Res. Commun., 363, 12, 10.1016/j.bbrc.2007.07.201 · DOI
  11. Green, 1981, Membrane phospholipid synthesis in Escherichia coli. Purification, reconstitution, and characterization of sn-glycerol-3-phosphate acyltransferase, J. Biol. Chem., 256, 11151, 10.1016/S0021-9258(19)68570-4 · DOI
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Metadata

Authors
Kuruma, Y., Stano, P., Ueda, T., & Luisi, P. L.
Venue
Biochimica et Biophysica Acta (BBA) - Biomembranes
Publisher
Elsevier BV
Published
2009-02
Volume
1788
Issue
2
Pages
567-574
ISSN
0005-2736 (ISSN-L)
OpenAlex
W1987445483
Cited by
248
References
4
FWCI
5.46
Type
journal article
Language
en
Open access
bronze
Licence

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CoChat. (2026). Citation check: A synthetic biology approach to the construction of membrane proteins in semi-synthetic minimal cells. https://app.cochat.ai/papers/10.1016/j.bbamem.2008.10.017
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Kuruma, Y., Stano, P., Ueda, T., & Luisi, P. L. (2009). A synthetic biology approach to the construction of membrane proteins in semi-synthetic minimal cells. Biochimica et Biophysica Acta (BBA) - Biomembranes. https://doi.org/10.1016/j.bbamem.2008.10.017

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