Signal peptidase I of Bacillus subtilis: patterns of conserved amino acids in prokaryotic and eukaryotic type I signal peptidases.

Dijl, J. M. van; Jong, Anne de; Vehmaanperä, Jari; Venema, Gerard; Bron, Sierd

doi:10.1002/j.1460-2075.1992.tb05349.x

Cited by 154 publications

(123 citation statements)

References 66 publications

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“…Current models predict that Implp and Imp2p have one amino terminal membrane spanning region ( Fig. 1A; Dalbey & von Heijne, 1992;van Dijl et al, 1992), although this has not been shown experimentally. In addition, Imp2p may have a carboxyl terminal membrane spanning region, as predicted for the signal peptidase of R. capsulutus (Fig.…”

Section: Mitochondrial Signal Peptidasementioning

confidence: 97%

“…1A). The latter feature is shared by the signal peptidase of the cyanobacterium Phonnidium laminosum (22 kDa; Packer et al, 1995), and all known signal peptidases from Gram positive bacteria, which include enzymes from Bacillus subtilis (van Dijl et al, 1992), Bacillus amyloliquefaciens, Bacillus culdolyticus, and Bacillus lichenifonnis (for a recent compilation of sequences see Meijer et al, 1995), Staphylococcus aureus (SpsB; Cregg et al, 1996), and Mycobacterium tuberculosis (Philipp et al, 1996). All Bacillus signal peptidases and the S. uureus signal peptidase consist of 21 kDa polypeptides whereas the M. tuberculosis gene encodes a 32 kDa polypeptide.…”

Section: Eubacterial Signal (Leader) Peptidasementioning

confidence: 99%

“…Interestingly, both in B. subtilis and B. amyloliquefaciens, chromosomal genes were identified for two homologous, but nonidentical, type I signal peptidases, denoted Sips and SipT (van Dijl et al, 1992;Meijer et al, 1995;Hoang & Hofemeister 1995;GenBank accession #U45883). In addition, a third chromosomal gene for a homologous type I signal peptidase (SipU) was identified in B. subtilis (Akagawa et al, 1995; H. Tjalsma, S. Bron & J. M. van Dijl, unpubl.…”

Section: Eubacterial Signal (Leader) Peptidasementioning

confidence: 99%

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The chemistry and enzymology of the type I signal peptidases

et al. 1997

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The discovery that proteins exported from the cytoplasm are typically synthesized as larger precursors with cleavable signal peptides has focused interest on the peptidases that remove the signal peptides. Here, we review the membranebound peptidases dedicated to the processing of protein precursors that are found in the plasma membrane of prokaryotes and the endoplasmic reticulum, the mitochondrial inner membrane, and the chloroplast thylakoidal membrane of eukaryotes. These peptidases are termed type I signal (or leader) peptidases. They share the unusual feature of being resistant to the general inhibitors of the four well-characterized peptidase classes. The eukaryotic and prokaryotic signal peptidases appear to belong to a single peptidase family. This review emphasizes the evolutionary concepts, current knowledge of the catalytic mechanism, and substrate specificity requirements of the signal peptidases.

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Section: Mitochondrial Signal Peptidasementioning

confidence: 97%

Section: Eubacterial Signal (Leader) Peptidasementioning

confidence: 99%

Section: Eubacterial Signal (Leader) Peptidasementioning

confidence: 99%

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The chemistry and enzymology of the type I signal peptidases

et al. 1997

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“…SPase I 1 has been isolated from Gram-negative (4 -6) and Gram-positive (7,8) bacteria as well as from several eukaryotic organisms (9 -12). These endopeptidases are membrane-bound and specific for the region within the signal peptide immediately preceding the cleavage site.…”

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confidence: 99%

The Role of the Conserved Box E Residues in the Active Site of the Escherichia coli Type I Signal Peptidase

Klenotic¹,

Carlos²,

Schuenemann³

et al. 2000

Journal of Biological Chemistry

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Type I signal peptidases are integral membrane proteins that function to remove signal peptides from secreted and membrane proteins. These enzymes carry out catalysis using a serine/lysine dyad instead of the prototypical serine/histidine/aspartic acid triad found in most serine proteases. Site-directed scanning mutagenesis was used to obtain a qualitative assessment of which residues in the fifth conserved region, Box E, of the Escherichia coli signal peptidase I are critical for maintaining a functional enzyme. First, we find that there is no requirement for activity for a salt bridge between the invariant Asp-273 and the Arg-146 residues. In addition, we show that the conserved Ser-278 is required for optimal activity as well as conserved salt bridge partners Asp-280 and Arg-282. Finally, Gly-272 is essential for signal peptidase I activity, consistent with it being located within van der Waals proximity to Ser-278 and general base Lys-145 side-chain atoms. We propose that replacement of the hydrogen side chain of Gly-272 with a methyl group results in steric crowding, perturbation of the active site conformation, and specifically, disruption of the Ser-90/Lys-145 hydrogen bond. A refined model is proposed for the catalytic dyad mechanism of signal peptidase I in which the general base Lys-145 is positioned by Ser-278, which in turn is held in place by Asp-280.The majority of proteins exported from a cell are made with a signal or leader peptide. These peptides are responsible for targeting proteins to the cytoplasmic membrane in prokaryotes and the endoplasmic reticulum membrane in eukaryotes (1-3). The protein is then translocated across the membrane, and the N-terminal peptide is removed from the pre-protein by a type I signal peptidase.SPase I 1 has been isolated from Gram-negative (4 -6) and Gram-positive (7, 8) bacteria as well as from several eukaryotic organisms (9 -12). These endopeptidases are membrane-bound and specific for the region within the signal peptide immediately preceding the cleavage site. The substrate protein is cleaved during or after the protein is transported across the membrane bilayer. Cleavage occurs by way of nucleophilic attack by a catalytic serine O␥ on the peptide bond between the presequence and the mature region of the pre-protein.The best-studied enzyme of this family is SPase I, or leader peptidase, of Escherichia coli. It has been cloned (13), sequenced (14), and purified (15, 16). Its substrate specificity has been characterized; small-uncharged residues at the P1 (Ϫ1) and P3 (Ϫ3) positions of the substrate are required for cleavage (17)(18)(19). SPase I utilizes a serine/lysine dyad mechanism to perform its enzymatic function rather than the canonical catalytic triad found in most serine proteases (20 -23). To date, only a few other enzymes, such as LexA (24), UmuD (25), and the Tsp protease (26) have been identified with this unusual active site mechanism. Additional analysis is required to provide insight into other residues near this dyad in the SPase family and th...

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“…Although the primary structure of signal peptides is poorly conserved, three functional domains have to be present: first, a positively charged amino terminus (N-region); second, a central hydrophobic domain (H-region); and third, a polar carboxyl-terminal domain (C-region), specifying the SPase cleavage site (1). We have previously shown that five paralogous type I SPases are involved in the processing of secretory precursor proteins in Bacillus subtilis (5)(6)(7). Two of these, denoted SipS and SipT, are of major importance for protein secretion.…”

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confidence: 99%

The Potential Active Site of the Lipoprotein-specific (Type II) Signal Peptidase of Bacillus subtilis

Tjalsma¹,

Zanen

Venema

et al. 1999

Journal of Biological Chemistry

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Type II signal peptidases (SPase II) remove signal peptides from lipid-modified preproteins of eubacteria. As the catalytic mechanism employed by type II SPases was not known, the present studies were aimed at the identification of their potential active site residues. Comparison of the deduced amino acid sequences of 19 known type II SPases revealed the presence of five conserved domains. The importance of the 15 best conserved residues in these domains was investigated using the type II SPase of Bacillus subtilis, which, unlike SPase II of Escherichia coli, is not essential for viability. The results showed that only six residues are important for SPase II activity. These are Asp-14, Asn-99, Asp-102, Asn-126, Ala-128, and Asp-129. Only Asp-14 was required for stability of SPase II, indicating that the other five residues are required for catalysis, the active site geometry, or the specific recognition of lipid-modified preproteins. As Asp-102 and Asp-129 are the only residues invoked in the known catalytic mechanisms of proteases, we hypothesize that these two residues are directly involved in SPase II-mediated catalysis. This implies that type II SPases belong to a novel family of aspartic proteases.

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Signal peptidase I of Bacillus subtilis: patterns of conserved amino acids in prokaryotic and eukaryotic type I signal peptidases.

Cited by 154 publications

References 66 publications

The chemistry and enzymology of the type I signal peptidases

The chemistry and enzymology of the type I signal peptidases

The Role of the Conserved Box E Residues in the Active Site of the Escherichia coli Type I Signal Peptidase

The Potential Active Site of the Lipoprotein-specific (Type II) Signal Peptidase of Bacillus subtilis

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