Institute of Molecular and Cellular Biosciences, The University of Tokyo,
(Present address)Cryobiofrontier Research Center, Faculty of Agriculture, Iwate University
Suntory Institute for Bioorganic Research
Department of Medical Genome Sciences, Graduate School of Frontier Sciences, The University of Tokyo
Department of Medical Genome Sciences, Graduate School of Frontier Sciences, The University of Tokyo
Suntory Institute for Bioorganic Research
Suntory Institute for Bioorganic Research
Department of Medical Genome Sciences, Graduate School of Frontier Sciences, The University of Tokyo
Institute of Molecular and Cellular Biosciences, The University of Tokyo
登録日
2010-10-05
雑誌名
Biochemical and Biophysical Research Communications
巻
394
号
3
ページ
733 - 736
発行年
2010-04-01
ISSN
0006-291X
Abstract
A complete reconstitution system for membrane integration of the simplest protein was developed by means of defined factors. A mutant version of Pf3 coat protein, 3L-Pf3 coat, requires neither signal recognition particle/Sec factors nor a membrane potential for its integration into the cytoplasmic membrane of Escherichia coli. Although 3L-Pf3 coat is spontaneously integrated into liposomes composed of phospholipids, diacylglycerol completely blocks such spontaneous integrations at a physiological level. Under the conditions where spontaneous integration does not occur, 3L-Pf3 coat integration was absolutely dependent on a novel integration-stimulating factor. Combination of the PURE system, an in vitro translation system composed of the purified factors involved in translation in E. coli, with liposomes containing the highly purified integration-stimulating factor revealed multiple cycles of 3L-Pf3 coat integration, achieving the complete reconstitution of membrane integration. Based on the function of the factor, we propose that the factor is named MPIase (Membrane Protein Integrase).
A novel complete reconstitution system for membrane integration of the simplest membrane protein
bbrc-v394n3p733-736.pdf
(2.52MB)
