Negatively charged liposome as a potent inhibitor of post-translation during in vitro synthesis of green fluorescent protein

Huong Thi Bui, Hiroshi Umakoshi, Keishi Suga, Masato Nishida, Toshinori Shimanouchi, Ryoichi Kuboi

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

The effect of negatively charged liposome on in vitro synthesis of a reporter protein, green fluorescent protein (GFP), was investigated using a cell-free translation system. GFP was expressed with and without negatively charged liposome prepared with 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylchorine (POPC) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoglycerol (POPG), resulting in the GFP fluorescence being reduced to approximately 60% in the presence of POPC/POPG at more than 30% POPG, depending on its concentration. However, the amount of synthesized GFP products, as analyzed by SDS-PAGE, did not change with and without the POPC/POPG liposome. The results of the ultrafiltration operation indicate that the liposome interacts not only with synthesized polypeptide GFP but also with folded synthesized GFP (mature GFP). Liposome also inhibited refolding of unfolded GFP to its native state. The above results show that the POPC/POPG could inhibit the folding of GFP in the post-translational process of the gene expression product.

Original languageEnglish
Pages (from-to)154-160
Number of pages7
JournalBiochemical Engineering Journal
Volume46
Issue number2
DOIs
Publication statusPublished - Oct 1 2009
Externally publishedYes

Fingerprint

Liposomes
Green Fluorescent Proteins
Proteins
In Vitro Techniques
Cell-Free System
Ultrafiltration
Polypeptides
Gene expression
Polyacrylamide Gel Electrophoresis
Fluorescence
Gene Expression
Peptides
1-palmitoyl-2-oleoylglycero-3-phosphoglycerol

Keywords

  • Green fluorescent protein (GFP)
  • Hydrophobicity
  • Liposome
  • Protein folding

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biomedical Engineering
  • Environmental Engineering

Cite this

Negatively charged liposome as a potent inhibitor of post-translation during in vitro synthesis of green fluorescent protein. / Bui, Huong Thi; Umakoshi, Hiroshi; Suga, Keishi; Nishida, Masato; Shimanouchi, Toshinori; Kuboi, Ryoichi.

In: Biochemical Engineering Journal, Vol. 46, No. 2, 01.10.2009, p. 154-160.

Research output: Contribution to journalArticle

Bui, Huong Thi ; Umakoshi, Hiroshi ; Suga, Keishi ; Nishida, Masato ; Shimanouchi, Toshinori ; Kuboi, Ryoichi. / Negatively charged liposome as a potent inhibitor of post-translation during in vitro synthesis of green fluorescent protein. In: Biochemical Engineering Journal. 2009 ; Vol. 46, No. 2. pp. 154-160.
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