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

Yutetsu Kuruma, Pasquale Stano, Takuya Ueda, Pier Luigi Luisi

Research output: Contribution to journalArticle

159 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)567-574
Number of pages8
JournalBiochimica et Biophysica Acta - Biomembranes
Volume1788
Issue number2
DOIs
Publication statusPublished - 2009 Jan 1

Keywords

  • Cell-free translation system
  • Liposome
  • Membrane protein
  • Minimal cell
  • Phospholipid biosynthesis
  • Self-reproduction

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Cell Biology

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