Flanking signal and mature peptide residues influence signal peptide cleavage

Choo, Khar Heng; Ranganathan, Shoba

doi:10.1186/1471-2105-9-s12-s15

Cited by 69 publications

(87 citation statements)

References 63 publications

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“…The secretions of the two lipases GehC and GehD also showed very different dose dependencies, with the former being more inhibited than the latter. This finding suggests that different Sec-type signal peptides are recognized by SPase with different affinities (although the origins of the differential recognition need not reside entirely within the signal peptide sequence itself [14]). This in turn suggests that under conditions of limiting SPase activity, SPase specificity may contribute a previously unappreciated level of control over the secretome.…”

Section: Discussionmentioning

confidence: 91%

“…While Ala is most common at both the Ϫ1 and Ϫ3 positions, Gly, Ser, Cys, and Thr are also found at the Ϫ1 position, and Gly, Ser, Cys, Ile, Val, and Leu are also found at the Ϫ3 position. The surrounding residues at the ϩ1, Ϫ2, Ϫ4, and Ϫ5 positions appear to be less critical, with most residues tolerated, with the exception of Pro at the ϩ1 position (42), and although a role for the residues beyond the ϩ1 position have been noted (14), their contribution is not well understood. Thus, while many trends have been identified, the prediction of signal peptides recognized by SPase based on sequence alone remains challenging and is further complicated by signal peptides that only poorly conform to the consensus rules (2,3,25,33,44,59).…”

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confidence: 99%

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Type I Signal Peptidase and Protein Secretion inStaphylococcus epidermidis

et al. 2011

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Bacterial protein secretion is a highly orchestrated process that is essential for infection and virulence. Despite extensive efforts to predict or experimentally detect proteins that are secreted, the characterization of the bacterial secretome has remained challenging. A central event in protein secretion is the type I signal peptidase (SPase)-mediated cleavage of the N-terminal signal peptide that targets a protein for secretion via the general secretory pathway, and the arylomycins are a class of natural products that inhibit SPase, suggesting that they may be useful chemical biology tools for characterizing the secretome. Here, using an arylomycin derivative, along with two-dimensional gel electrophoresis and liquid chromatography-tandem mass spectrometry (LC-MS/MS), we identify 11 proteins whose secretion from stationary-phase Staphylococcus epidermidis is dependent on SPase activity, 9 of which are predicted to be translated with canonical N-terminal signal peptides. In addition, we find that the presence of extracellular domains of lipoteichoic acid synthase (LtaS) and the ␤-lactam response sensor BlaR1 in the medium is dependent on SPase activity, suggesting that they are cleaved at noncanonical sites within the protein. In all, the data define the proteins whose stationaryphase secretion depends on SPase and also suggest that the arylomycins should be valuable chemical biology tools for the study of protein secretion in a wide variety of different bacteria.

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Section: Discussionmentioning

confidence: 91%

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confidence: 99%

Type I Signal Peptidase and Protein Secretion inStaphylococcus epidermidis

et al. 2011

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“…They interact with the SecA protein, the signal recognition particle (SRP), and the signal peptidase (16,30). The interaction between the SP and the mature protein is known to influence protein export as well (9,16,17). Therefore, the choice of an efficient signal peptide for any given target protein is of utmost importance, and several approaches to identify efficient SPs for different target proteins were taken (2,4,6,15,21,38).…”

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confidence: 99%

Optimization of Protease Secretion in Bacillus subtilis and Bacillus licheniformis by Screening of Homologous and Heterologous Signal Peptides

Degering

Eggert²,

Puls³

et al. 2010

Appl Environ Microbiol

163

117

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Bacillus subtilis and Bacillus licheniformis are widely used for the large-scale industrial production of proteins. These strains can efficiently secrete proteins into the culture medium using the general secretion (Sec) pathway. A characteristic feature of all secreted proteins is their N-terminal signal peptides, which are recognized by the secretion machinery. Here, we have studied the production of an industrially important secreted protease, namely, subtilisin BPN from Bacillus amyloliquefaciens. One hundred seventy-three signal peptides originating from B. subtilis and 220 signal peptides from the B. licheniformis type strain were fused to this secretion target and expressed in B. subtilis, and the resulting library was analyzed by high-throughput screening for extracellular proteolytic activity. We have identified a number of signal peptides originating from both organisms which produced significantly increased yield of the secreted protease. Interestingly, we observed that levels of extracellular protease were improved not only in B. subtilis, which was used as the screening host, but also in two different B. licheniformis strains. To date, it is impossible to predict which signal peptide will result in better secretion and thus an improved yield of a given extracellular target protein. Our data show that screening a library consisting of homologous and heterologous signal peptides fused to a target protein can identify more-effective signal peptides, resulting in improved protein export not only in the original screening host but also in different production strains.Gram-positive bacteria of the genus Bacillus are industrially well-established microorganisms for the production of extracellular proteins. Due to the availability of relatively cheap large-scale production systems combined with the ability of bacteria to secrete up to 20 to 25 g/liter of a target protein into the growth medium, about 60% of commercially available enzymes are presently produced in Bacillus species (14, 28).The closely related species Bacillus subtilis and Bacillus licheniformis are widely used as production hosts on an industrial scale, and, in contrast to the well-known production species Escherichia coli, they are free of endotoxin and have GRAS (generally regarded as safe) status. The complete genome sequences of strains B. subtilis 168 (1, 18) and B. licheniformis DSM13 (isogenic to ATCC 14580) (26, 32) are available, greatly facilitating the construction of improved production strains.The Sec pathway constitutes the main secretion pathway in B. subtilis and B. licheniformis. Proteins secreted via the Sec pathway are initially synthesized with an N-terminal hydrophobic signal peptide (SP) consisting of a positively charged N domain followed by a longer, hydrophobic H domain and a C domain consisting of three amino acids which form the signal peptidase recognition site (35). Targeting of a secreted protein to the membrane, the translocation process itself, and subsequent processing by a signal peptidase represent the majo...

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“…Although there is no canonical motif in SPs that predicts cleavage by signal peptidases, SP prediction programs can identify potential SP cleavage sites with high confidence via a combination of contextual cues (refer- ence 13 and references therein). The median length of eukaryotic signal peptides is 22 amino acids (aa) (range, 11 to 55 aa) (12), and SPs were clearly present in the first ϳ25 amino acids of the F proteins from NiV Malaysia , NiV Bangladesh , HeV, and CedPV but not in GhV-F= (Fig. 3B).…”

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confidence: 99%

“…Signal peptides (SPs) are usually located within the first 20 to 30 amino acids of the N termini of plasma membrane-associated or secreted proteins, and they are characterized by a preponderance of aliphatic and hydrophobic amino acids, such as leucines and isoleucines (12). Signal peptidases cleave SPs to generate the mature forms of the associated proteins.…”

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confidence: 99%

Functional Rectification of the Newly Described African Henipavirus Fusion Glycoprotein (Gh-M74a)

Pernet

Beaty

Lee

2014

J Virol

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bRecent evidence identified multiple Henipavirus species in Africa distinct from those in Southeast Asia and Australia. The reported fusion glycoprotein (F) sequence of the African Gh-M74a strain (GhV-F) is likely incorrect: a single base pair deletion near the N terminus results in multiple aberrancies. Rectifying this by adding single nucleotide insertions results in a GhV-F that now possesses a signal peptide, is efficiently cell surface expressed, exhibits syncytium formation when coexpressed with GhV-G protein, and mediates pseudotyped viral particle entry.

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Flanking signal and mature peptide residues influence signal peptide cleavage

Cited by 69 publications

References 63 publications

Type I Signal Peptidase and Protein Secretion inStaphylococcus epidermidis

Type I Signal Peptidase and Protein Secretion inStaphylococcus epidermidis

Optimization of Protease Secretion in Bacillus subtilis and Bacillus licheniformis by Screening of Homologous and Heterologous Signal Peptides

Functional Rectification of the Newly Described African Henipavirus Fusion Glycoprotein (Gh-M74a)

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