TY - JOUR
T1 - A novel complete reconstitution system for membrane integration of the simplest membrane protein
AU - Nishiyama, Ken ichi
AU - Maeda, Masahide
AU - Abe, Masato
AU - Kanamori, Takashi
AU - Shimamoto, Keiko
AU - Kusumoto, Shoichi
AU - Ueda, Takuya
AU - Tokuda, Hajime
N1 - Funding Information:
We thank Prof. Andreas Kuhn for the gift of the plasmid. This work was supported by grants-in-aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan .
PY - 2010/4/9
Y1 - 2010/4/9
N2 - 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).
AB - 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).
KW - Diacylglycerol
KW - Integration-stimulating factor
KW - Liposomes
KW - MPIase
KW - Membrane protein integration
KW - PURE system
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U2 - 10.1016/j.bbrc.2010.03.061
DO - 10.1016/j.bbrc.2010.03.061
M3 - Article
C2 - 20230783
AN - SCOPUS:77950518678
SN - 0006-291X
VL - 394
SP - 733
EP - 736
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 3
ER -