Signal peptide hydrophobicity is critical for early stages in protein export by Bacillus subtilis

L.F. van Zanen, E.N.G. Houben, R.B. Meima, H. Tjalsma, J.D.H. Jongbloed, T.M. Westers, B. Oudega, S. Luirink, J.M. Dijl, W.J. Quax

Research output: Contribution to JournalArticleAcademicpeer-review

Abstract

Signal peptides that direct protein export in Bacillus subtilis are overall more hydrophobic than signal peptides in Escherichia coli. To study the importance of signal peptide hydrophobicity for protein export in both organisms, the α-amylase AmyQ was provided with leucine-rich (high hydrophobicity) or alanine-rich (low hydrophobicity) signal peptides. AmyQ export was most efficiently directed by the authentic signal peptide, both in E. coli and B. subtilis. The leucine-rich signal peptide directed AmyQ export less efficiently in both organisms, as judged from pulse-chase labelling experiments. Remarkably, the alanine-rich signal peptide was functional in protein translocation only in E. coli. Cross-linking of in vitro synthesized ribosome nascent chain complexes (RNCs) to cytoplasmic proteins showed that signal peptide hydrophobicity is a critical determinant for signal peptide binding to the Ffh component of the signal recognition particle (SRP) or to trigger factor, not only in E. coli, but also in B. subtilis. The results show that B. subtilis SRP can discriminate between signal peptides with relatively high hydrophobicities. Interestingly, the B. subtilis protein export machinery seems to be poorly adapted to handle alanine-rich signal peptides with a low hydrophobicity. Thus, signal peptide hydrophobicity appears to be more critical for the efficiency of early stages in protein export in B. subtilis than in E. coli. © 2005 FEBS.
Original languageEnglish
Pages (from-to)4617-4630
Number of pages14
JournalThe FEBS Journal
Volume272
DOIs
Publication statusPublished - 2005

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Bacilli
Hydrophobicity
Protein Sorting Signals
Bacillus subtilis
Hydrophobic and Hydrophilic Interactions
Proteins
Escherichia coli
Signal Recognition Particle
Alanine
Leucine
Protein Transport
Amylases
Ribosomes
Labeling
Machinery

Cite this

van Zanen, L. F., Houben, E. N. G., Meima, R. B., Tjalsma, H., Jongbloed, J. D. H., Westers, T. M., ... Quax, W. J. (2005). Signal peptide hydrophobicity is critical for early stages in protein export by Bacillus subtilis. The FEBS Journal, 272, 4617-4630. https://doi.org/10.1111/j.1742-4658.2005.04777.x
van Zanen, L.F. ; Houben, E.N.G. ; Meima, R.B. ; Tjalsma, H. ; Jongbloed, J.D.H. ; Westers, T.M. ; Oudega, B. ; Luirink, S. ; Dijl, J.M. ; Quax, W.J. / Signal peptide hydrophobicity is critical for early stages in protein export by Bacillus subtilis. In: The FEBS Journal. 2005 ; Vol. 272. pp. 4617-4630.
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abstract = "Signal peptides that direct protein export in Bacillus subtilis are overall more hydrophobic than signal peptides in Escherichia coli. To study the importance of signal peptide hydrophobicity for protein export in both organisms, the α-amylase AmyQ was provided with leucine-rich (high hydrophobicity) or alanine-rich (low hydrophobicity) signal peptides. AmyQ export was most efficiently directed by the authentic signal peptide, both in E. coli and B. subtilis. The leucine-rich signal peptide directed AmyQ export less efficiently in both organisms, as judged from pulse-chase labelling experiments. Remarkably, the alanine-rich signal peptide was functional in protein translocation only in E. coli. Cross-linking of in vitro synthesized ribosome nascent chain complexes (RNCs) to cytoplasmic proteins showed that signal peptide hydrophobicity is a critical determinant for signal peptide binding to the Ffh component of the signal recognition particle (SRP) or to trigger factor, not only in E. coli, but also in B. subtilis. The results show that B. subtilis SRP can discriminate between signal peptides with relatively high hydrophobicities. Interestingly, the B. subtilis protein export machinery seems to be poorly adapted to handle alanine-rich signal peptides with a low hydrophobicity. Thus, signal peptide hydrophobicity appears to be more critical for the efficiency of early stages in protein export in B. subtilis than in E. coli. {\circledC} 2005 FEBS.",
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Signal peptide hydrophobicity is critical for early stages in protein export by Bacillus subtilis. / van Zanen, L.F.; Houben, E.N.G.; Meima, R.B.; Tjalsma, H.; Jongbloed, J.D.H.; Westers, T.M.; Oudega, B.; Luirink, S.; Dijl, J.M.; Quax, W.J.

In: The FEBS Journal, Vol. 272, 2005, p. 4617-4630.

Research output: Contribution to JournalArticleAcademicpeer-review

TY - JOUR

T1 - Signal peptide hydrophobicity is critical for early stages in protein export by Bacillus subtilis

AU - van Zanen, L.F.

AU - Houben, E.N.G.

AU - Meima, R.B.

AU - Tjalsma, H.

AU - Jongbloed, J.D.H.

AU - Westers, T.M.

AU - Oudega, B.

AU - Luirink, S.

AU - Dijl, J.M.

AU - Quax, W.J.

PY - 2005

Y1 - 2005

N2 - Signal peptides that direct protein export in Bacillus subtilis are overall more hydrophobic than signal peptides in Escherichia coli. To study the importance of signal peptide hydrophobicity for protein export in both organisms, the α-amylase AmyQ was provided with leucine-rich (high hydrophobicity) or alanine-rich (low hydrophobicity) signal peptides. AmyQ export was most efficiently directed by the authentic signal peptide, both in E. coli and B. subtilis. The leucine-rich signal peptide directed AmyQ export less efficiently in both organisms, as judged from pulse-chase labelling experiments. Remarkably, the alanine-rich signal peptide was functional in protein translocation only in E. coli. Cross-linking of in vitro synthesized ribosome nascent chain complexes (RNCs) to cytoplasmic proteins showed that signal peptide hydrophobicity is a critical determinant for signal peptide binding to the Ffh component of the signal recognition particle (SRP) or to trigger factor, not only in E. coli, but also in B. subtilis. The results show that B. subtilis SRP can discriminate between signal peptides with relatively high hydrophobicities. Interestingly, the B. subtilis protein export machinery seems to be poorly adapted to handle alanine-rich signal peptides with a low hydrophobicity. Thus, signal peptide hydrophobicity appears to be more critical for the efficiency of early stages in protein export in B. subtilis than in E. coli. © 2005 FEBS.

AB - Signal peptides that direct protein export in Bacillus subtilis are overall more hydrophobic than signal peptides in Escherichia coli. To study the importance of signal peptide hydrophobicity for protein export in both organisms, the α-amylase AmyQ was provided with leucine-rich (high hydrophobicity) or alanine-rich (low hydrophobicity) signal peptides. AmyQ export was most efficiently directed by the authentic signal peptide, both in E. coli and B. subtilis. The leucine-rich signal peptide directed AmyQ export less efficiently in both organisms, as judged from pulse-chase labelling experiments. Remarkably, the alanine-rich signal peptide was functional in protein translocation only in E. coli. Cross-linking of in vitro synthesized ribosome nascent chain complexes (RNCs) to cytoplasmic proteins showed that signal peptide hydrophobicity is a critical determinant for signal peptide binding to the Ffh component of the signal recognition particle (SRP) or to trigger factor, not only in E. coli, but also in B. subtilis. The results show that B. subtilis SRP can discriminate between signal peptides with relatively high hydrophobicities. Interestingly, the B. subtilis protein export machinery seems to be poorly adapted to handle alanine-rich signal peptides with a low hydrophobicity. Thus, signal peptide hydrophobicity appears to be more critical for the efficiency of early stages in protein export in B. subtilis than in E. coli. © 2005 FEBS.

U2 - 10.1111/j.1742-4658.2005.04777.x

DO - 10.1111/j.1742-4658.2005.04777.x

M3 - Article

VL - 272

SP - 4617

EP - 4630

JO - The FEBS Journal

JF - The FEBS Journal

SN - 1742-464X

ER -