Consequences of the overexpression of a eukaryotic membrane protein, the human KDEL receptor, in Escherichia coli
Standard
Consequences of the overexpression of a eukaryotic membrane protein, the human KDEL receptor, in Escherichia coli. / Klepsch, Mirjam M; Persson, Jan O; de Gier, Jan-Willem L.
In: J MOL BIOL, Vol. 407, No. 4, 08.04.2011, p. 532-42.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
Harvard
APA
Vancouver
Bibtex
}
RIS
TY - JOUR
T1 - Consequences of the overexpression of a eukaryotic membrane protein, the human KDEL receptor, in Escherichia coli
AU - Klepsch, Mirjam M
AU - Persson, Jan O
AU - de Gier, Jan-Willem L
N1 - Copyright © 2011 Elsevier Ltd. All rights reserved.
PY - 2011/4/8
Y1 - 2011/4/8
N2 - Escherichia coli is the most widely used host for producing membrane proteins. Thus far, to study the consequences of membrane protein overexpression in E. coli, we have focussed on prokaryotic membrane proteins as overexpression targets. Their overexpression results in the saturation of the Sec translocon, which is a protein-conducting channel in the cytoplasmic membrane that mediates both protein translocation and insertion. Saturation of the Sec translocon leads to (i) protein misfolding/aggregation in the cytoplasm, (ii) impaired respiration, and (iii) activation of the Arc response, which leads to inefficient ATP production and the formation of acetate. The overexpression yields of eukaryotic membrane proteins in E. coli are usually much lower than those of prokaryotic ones. This may be due to differences between the consequences of the overexpression of prokaryotic and eukaryotic membrane proteins in E. coli. Therefore, we have now also studied in detail how the overexpression of a eukaryotic membrane protein, the human KDEL receptor, affects E. coli. Surprisingly, the consequences of the overexpression of a prokaryotic and a eukaryotic membrane protein are very similar. Strain engineering and likely also protein engineering can be used to remedy the saturation of the Sec translocon upon overexpression of both prokaryotic and eukaryotic membrane proteins in E. coli.
AB - Escherichia coli is the most widely used host for producing membrane proteins. Thus far, to study the consequences of membrane protein overexpression in E. coli, we have focussed on prokaryotic membrane proteins as overexpression targets. Their overexpression results in the saturation of the Sec translocon, which is a protein-conducting channel in the cytoplasmic membrane that mediates both protein translocation and insertion. Saturation of the Sec translocon leads to (i) protein misfolding/aggregation in the cytoplasm, (ii) impaired respiration, and (iii) activation of the Arc response, which leads to inefficient ATP production and the formation of acetate. The overexpression yields of eukaryotic membrane proteins in E. coli are usually much lower than those of prokaryotic ones. This may be due to differences between the consequences of the overexpression of prokaryotic and eukaryotic membrane proteins in E. coli. Therefore, we have now also studied in detail how the overexpression of a eukaryotic membrane protein, the human KDEL receptor, affects E. coli. Surprisingly, the consequences of the overexpression of a prokaryotic and a eukaryotic membrane protein are very similar. Strain engineering and likely also protein engineering can be used to remedy the saturation of the Sec translocon upon overexpression of both prokaryotic and eukaryotic membrane proteins in E. coli.
KW - Acetates
KW - Adenosine Triphosphatases
KW - Bacterial Proteins
KW - Escherichia coli
KW - Gene Expression
KW - Humans
KW - Membrane Proteins
KW - Membrane Transport Proteins
KW - Protein Engineering
KW - Receptors, Peptide
KW - Recombinant Proteins
KW - SEC Translocation Channels
KW - Journal Article
KW - Research Support, N.I.H., Extramural
U2 - 10.1016/j.jmb.2011.02.007
DO - 10.1016/j.jmb.2011.02.007
M3 - SCORING: Journal article
C2 - 21316372
VL - 407
SP - 532
EP - 542
JO - J MOL BIOL
JF - J MOL BIOL
SN - 0022-2836
IS - 4
ER -