Effect of alteration of charged residues at the N termini of signal peptides on protein export in Bacillus subtilis

Chen, Mario; Nagarajan, Vasantha

doi:10.1128/jb.176.18.5796-5801.1994

Cited by 28 publications

(19 citation statements)

References 32 publications

(28 reference statements)

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“…19). A systematic study of the influence of signal peptide variations on the secretion of Bacillus amyloliquefaciens levansucrase in Bacillus subtilis (20,21) is in general agreement with the results obtained for E. coli. However, the dependence on positive charges in the N-terminal region was more pronounced; signal peptides with neutral net charge significantly impaired the export of the precusor protein.…”

supporting

confidence: 86%

Influence of Specific Signal Peptide Mutations on the Expression and Secretion of the α-Amylase Inhibitor Tendamistat in

Fass

Engels

1996

Journal of Biological Chemistry

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The Streptomyces ␣-amylase inhibitor tendamistat is secreted by a signal peptide with an amino-terminal charge of ؉3. To elucidate the influence of the charged residues on protein secretion in Streptomyces, the amino-terminal charge was varied from ؉6 to neutral net charge. The effects of charge variation were analyzed in combination with three Streptomyces promoters and two transcriptional terminators. Introduction of additional positive charges significantly decreased the amount of secreted tendamistat. On the contrary, a charge reduction to ؉2 resulted in the doubling of inhibitor production. After exclusion of transcriptional effects, the observed alterations of inhibitor secretion by the mutants with a charge of ؉6 to ؉2 were attributed to a modulation of precursor synthesis. Furthermore, a tight coupling of synthesis and export was stated. Charge reduction to ؉1 or neutral charge generally reduced the yield of secreted tendamistat, yet remarkable differences were found for mutants with identical net charge. Elimination of the positive charge at a defined position resulted in the release of tendamistat precursor protein, which suggested a specific uncoupling of synthesis and translocation.

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supporting

confidence: 86%

Influence of Specific Signal Peptide Mutations on the Expression and Secretion of the α-Amylase Inhibitor Tendamistat in

Fass

Engels

1996

Journal of Biological Chemistry

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“…In general, a signal sequence requires three distinct domains for its function (Pugsley, 1993;Tjalsma et al, 2000). The region adjacent to the N terminus, called the N-domain, contains several positively charged amino acids for attachment to the cell membrane (Akita et al, 1990;Chen & Nagarajan, 1994;Gennity et al, 1990). Next to the N-domain is the hydrophobic H-domain region, which forms an a-helical conformation in the membrane (Briggs et al, 1986).…”

Section: Introductionmentioning

confidence: 99%

Scanning the Corynebacterium glutamicum R genome for high-efficiency secretion signal sequences

et al. 2009

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Systematic screening of secretion proteins using an approach based on the completely sequenced genome of Corynebacterium glutamicum R revealed 405 candidate signal peptides, 108 of which were able to heterologously secrete an active-form a-amylase derived from Geobacillus stearothermophilus. These comprised 90 general secretory (Sec)-type, 10 twinarginine translocator (Tat)-type and eight Sec-type with presumptive lipobox peptides. Only Secand Tat-type signals directed high-efficiency secretion. In two assays, 11 of these signals resulted in 50-to 150-fold increased amounts of secreted a-amylase compared with the well-known corynebacterial secretory protein PS2. While the presence of an AXA motif at the cleavage sites was readily apparent, it was the presence of a glutamine residue adjacent to the cleavage site that may affect secretion efficiency.

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“…The aminoterminal N-domain of signal peptides contains at least one arginine or lysine residue, although this positively charged residue does not seem to be strictly required for protein export (51,101). The positively charged N-domain has been suggested to interact with the translocation machinery (1) and negatively charged phospholipids in the lipid bilayer of the membrane during translocation (71).…”

Section: Amino-terminal Signal Peptidesmentioning

confidence: 99%

Signal Peptide-Dependent Protein Transport in Bacillus subtilis : a Genome-Based Survey of the Secretome

Tjalsma

Bolhuis

Jongbloed

et al. 2000

Microbiol Mol Biol Rev

748

768

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SUMMARY One of the most salient features of Bacillus subtilis and related bacilli is their natural capacity to secrete a variety of proteins into their environment, frequently to high concentrations. This has led to the commercial exploitation of bacilli as major “cell factories” for secreted enzymes. The recent sequencing of the genome of B. subtilis has provided major new impulse for analysis of the molecular mechanisms underlying protein secretion by this organism. Most importantly, the genome sequence has allowed predictions about the composition of the secretome, which includes both the pathways for protein transport and the secreted proteins. The present survey of the secretome describes four distinct pathways for protein export from the cytoplasm and approximately 300 proteins with the potential to be exported. By far the largest number of exported proteins are predicted to follow the major “Sec” pathway for protein secretion. In contrast, the twin-arginine translocation “Tat” pathway, a type IV prepilin-like export pathway for competence development, and ATP-binding cassette transporters can be regarded as “special-purpose” pathways, through which only a few proteins are transported. The properties of distinct classes of amino-terminal signal peptides, directing proteins into the various protein transport pathways, as well as the major components of each pathway are discussed. The predictions and comparisons in this review pinpoint important differences as well as similarities between protein transport systems in B. subtilis and other well-studied organisms, such as Escherichia coli and the yeast Saccharomyces cerevisiae. Thus, they may serve as a lead for future research and applications.

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Effect of alteration of charged residues at the N termini of signal peptides on protein export in Bacillus subtilis

Cited by 28 publications

References 32 publications

Influence of Specific Signal Peptide Mutations on the Expression and Secretion of the α-Amylase Inhibitor Tendamistat in

Influence of Specific Signal Peptide Mutations on the Expression and Secretion of the α-Amylase Inhibitor Tendamistat in

Scanning the Corynebacterium glutamicum R genome for high-efficiency secretion signal sequences

Signal Peptide-Dependent Protein Transport in Bacillus subtilis : a Genome-Based Survey of the Secretome

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