Details
| Original language | English |
|---|---|
| Pages (from-to) | 329-336 |
| Number of pages | 8 |
| Journal | FEMS Microbiology Letters |
| Volume | 164 |
| Issue number | 2 |
| Publication status | Published - 15 Jul 1998 |
| Externally published | Yes |
Abstract
Proteins which are synthesized with a signal peptide containing a 'double-arginine' motif may be translocated across the bacterial cytoplasmic membrane by a mechanism that is different from the known Sec and signal recognition particle pathways. The function of the double-arginine motif as a determinant for this novel pathway was studied by expressions of gene constructs coding for the high potential iron-sulfur protein (HiPIP) from Chromatium vinosum D in Escherichia coli. When the protein was produced with its original double-arginine motif-containing signal peptide, it was in part translocated into the periplasm and thereby processed, as shown by immunoblots after cell fractionation and N-terminal sequencing of purified HiPIP. Processing was not inhibited significantly by 3 mM sodium azide, indicating that translocation of HiPIP occurs by a SecA-independent pathway. Translocation of HiPIP could be altered to the SecA-dependent mode when its signal peptide was substituted by that of PelB from Erwinia carotovora. When the HiPIP double-arginine motif (SRRDAVK) was introduced into the corresponding position of the PelB signal peptide, the transport pathway remained SecA-dependent. This indicates that additional determinants are required for translocation by the Sec-independent pathway. Copyright (C) 1998 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.
Keywords
- Chromatium vinosum, Double-arginine motif signal peptide, Escherichia coli, High potential iron-sulfur protein, Iron-sulfur cluster, Protein translocation
ASJC Scopus subject areas
- Immunology and Microbiology(all)
- Microbiology
- Biochemistry, Genetics and Molecular Biology(all)
- Molecular Biology
- Biochemistry, Genetics and Molecular Biology(all)
- Genetics
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In: FEMS Microbiology Letters, Vol. 164, No. 2, 15.07.1998, p. 329-336.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Evidence against the double-arginine motif as the only determinant for protein translocation by a novel Sec-independent pathway in Escherichia coli
AU - Brüser, Thomas
AU - Deutzmann, Rainer
AU - Dahl, Christiane
N1 - Funding Information: We thank H.-G. Sahl and M. Becker (Bonn, Germany) for production of antiserum. We also thank W. Klipp (Bochum, Germany), A. Seidler (Bochum, Germany) and J. Packer (Cambridge, England) for helpful comments and discussions. We are grateful to H.G. Trüper for his support of the project. T.B. thanks the Cusanuswerk for a grant. Copyright: Copyright 2007 Elsevier B.V., All rights reserved.
PY - 1998/7/15
Y1 - 1998/7/15
N2 - Proteins which are synthesized with a signal peptide containing a 'double-arginine' motif may be translocated across the bacterial cytoplasmic membrane by a mechanism that is different from the known Sec and signal recognition particle pathways. The function of the double-arginine motif as a determinant for this novel pathway was studied by expressions of gene constructs coding for the high potential iron-sulfur protein (HiPIP) from Chromatium vinosum D in Escherichia coli. When the protein was produced with its original double-arginine motif-containing signal peptide, it was in part translocated into the periplasm and thereby processed, as shown by immunoblots after cell fractionation and N-terminal sequencing of purified HiPIP. Processing was not inhibited significantly by 3 mM sodium azide, indicating that translocation of HiPIP occurs by a SecA-independent pathway. Translocation of HiPIP could be altered to the SecA-dependent mode when its signal peptide was substituted by that of PelB from Erwinia carotovora. When the HiPIP double-arginine motif (SRRDAVK) was introduced into the corresponding position of the PelB signal peptide, the transport pathway remained SecA-dependent. This indicates that additional determinants are required for translocation by the Sec-independent pathway. Copyright (C) 1998 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.
AB - Proteins which are synthesized with a signal peptide containing a 'double-arginine' motif may be translocated across the bacterial cytoplasmic membrane by a mechanism that is different from the known Sec and signal recognition particle pathways. The function of the double-arginine motif as a determinant for this novel pathway was studied by expressions of gene constructs coding for the high potential iron-sulfur protein (HiPIP) from Chromatium vinosum D in Escherichia coli. When the protein was produced with its original double-arginine motif-containing signal peptide, it was in part translocated into the periplasm and thereby processed, as shown by immunoblots after cell fractionation and N-terminal sequencing of purified HiPIP. Processing was not inhibited significantly by 3 mM sodium azide, indicating that translocation of HiPIP occurs by a SecA-independent pathway. Translocation of HiPIP could be altered to the SecA-dependent mode when its signal peptide was substituted by that of PelB from Erwinia carotovora. When the HiPIP double-arginine motif (SRRDAVK) was introduced into the corresponding position of the PelB signal peptide, the transport pathway remained SecA-dependent. This indicates that additional determinants are required for translocation by the Sec-independent pathway. Copyright (C) 1998 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.
KW - Chromatium vinosum
KW - Double-arginine motif signal peptide
KW - Escherichia coli
KW - High potential iron-sulfur protein
KW - Iron-sulfur cluster
KW - Protein translocation
UR - http://www.scopus.com/inward/record.url?scp=0039351902&partnerID=8YFLogxK
U2 - 10.1016/S0378-1097(98)00233-X
DO - 10.1016/S0378-1097(98)00233-X
M3 - Article
C2 - 9682482
AN - SCOPUS:0039351902
VL - 164
SP - 329
EP - 336
JO - FEMS Microbiology Letters
JF - FEMS Microbiology Letters
SN - 0378-1097
IS - 2
ER -