Crystal Structure of a Bacterial Signal Peptide Peptidase

Kim, Apollos C.; Oliver, David C.; Paetzel, Mark

doi:10.1016/j.jmb.2007.11.080

Cited by 45 publications

(57 citation statements)

References 39 publications

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“…Lys and Arg are also common in the interface regions of typical monotopic membrane proteins, e.g. prostaglandin H2 synthase-1 (56), signal peptide peptidase (61), and glycerol-3-phosphate dehydrogenase (62). Of the three monotopic proteins here with very similar three-dimensional structures (i.e.…”

Section: Formation Of Intracellular Membranesmentioning

confidence: 83%

Massive Formation of Intracellular Membrane Vesicles in Escherichia coli by a Monotopic Membrane-bound Lipid Glycosyltransferase

Eriksson

Wessman

et al. 2009

Journal of Biological Chemistry

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The morphology and curvature of biological bilayers are determined by the packing shapes and interactions of their participant molecules. Bacteria, except photosynthetic groups, usually lack intracellular membrane organelles. Strong overexpression in Escherichia coli of a foreign monotopic glycosyltransferase (named monoglycosyldiacylglycerol synthase), synthesizing a nonbilayer-prone glucolipid, induced massive formation of membrane vesicles in the cytoplasm. Vesicle assemblies were visualized in cytoplasmic zones by fluorescence microscopy. These have a very low buoyant density, substantially different from inner membranes, with a lipid content of >60% (w/w). Cryo-transmission electron microscopy revealed cells to be filled with membrane vesicles of various sizes and shapes, which when released were mostly spherical (diameter ≈100 nm). The protein repertoire was similar in vesicle and inner membranes and dominated by the glycosyltransferase. Membrane polar lipid composition was similar too, including the foreign glucolipid. A related glycosyltransferase and an inactive monoglycosyldiacylglycerol synthase mutant also yielded membrane vesicles, but without glucolipid synthesis, strongly indicating that vesiculation is induced by the protein itself. The high capacity for membrane vesicle formation seems inherent in the glycosyltransferase structure, and it depends on the following: (i) lateral expansion of the inner monolayer by interface binding of many molecules; (ii) membrane expansion through stimulation of phospholipid synthesis, by electrostatic binding and sequestration of anionic lipids; (iii) bilayer bending by the packing shape of excess nonbilayer-prone phospholipid or glucolipid; and (iv) potentially also the shape or penetration profile of the glycosyltransferase binding surface. These features seem to apply to several other proteins able to achieve an analogous membrane expansion.

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Section: Formation Of Intracellular Membranesmentioning

confidence: 83%

Massive Formation of Intracellular Membrane Vesicles in Escherichia coli by a Monotopic Membrane-bound Lipid Glycosyltransferase

Eriksson

Wessman

et al. 2009

Journal of Biological Chemistry

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“…Although SppA is generally considered to be the SP-cleaving enzyme in bacteria, disruption of the E. coli sppA gene did not affect in vivo cleavage of SPs of fusion proteins, including that from Lpp in our experiments. SppA has a large periplasmic domain, which, in archaea, forms a tetra- meric assembly of an inverted bowl-like shape, the membranedistal part of which includes the protease active sites (24). The possibility remains that SppA provides a route of SP degradation if SPs are released to the periplasm, although its in vivo role has not been studied.…”

Section: Discussionmentioning

confidence: 99%

Post-liberation cleavage of signal peptides is catalyzed by the site-2 protease (S2P) in bacteria

Saito

Hizukuri

Matsuo

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

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A signal peptide (SP) is cleaved off from presecretory proteins by signal peptidase during or immediately after insertion into the membrane. In metazoan cells, the cleaved SP then receives proteolysis by signal peptide peptidase, an intramembrane-cleaving protease (I-CLiP). However, bacteria lack any signal peptide peptidase member I-CLiP, and little is known about the metabolic fate of bacterial SPs. Here we show that Escherichia coli RseP, an site-2 protease (S2P) family I-CLiP, introduces a cleavage into SPs after their signal peptidase-mediated liberation from preproteins. A Bacillus subtilis S2P protease, RasP, is also shown to be involved in SP cleavage. These results uncover a physiological role of bacterial S2P proteases and update the basic knowledge about the fate of signal peptides in bacterial cells. SPP belongs to intramembrane-cleaving proteases (I-CLiPs), which are classified into SPP/γ-secretase (aspartyl proteases), rhomboid (serine protease), and S2P (zinc metalloprotease) (5, 6). These proteases liberate otherwise membrane-tethered domains of membrane proteins to function as a regulatory molecule. I-CLiPs have specific intramolecular routes that make water molecules accessible to the intramembrane proteolytic active sites (5, 6). For such proteolysis to occur, γ-secretase/SPP and S2P require, in most cases, removal of the ectodomain of the substrate by other protease (5, 7). SPP and S2P prefer substrate with the type II transmembrane orientation and γ-secretase and rhomboid prefer the type I orientation (7).Bacteria contain S2Ps and rhomboids but not SPPs, and only limited knowledge is available about the fate of bacterial SPs (7). We and others have been characterizing RseP, an Escherichia coli member of the S2P involved in the σ E pathway extracytoplasmic stress response (8, 9). In this regulation, a protease, DegS, responds to misassembled outer membrane proteins and introduces the first proteolytic cleavage into RseA, a membraneintegrated anti-σ E protein (10). Subsequently, RseP introduces the second cleavage into RseA, activating σ E to transcribe stressinducible genes (8, 9). Although RseA is the only physiological substrate of RseP so far established, we have shown previously that RseP can cleave a wider range of transmembrane sequences having helix-destabilizing residues (11). Our preliminary results that RseP cleaved a β-lactamase (Bla) fusion protein at or around its SP (11), together with the fact that RseP and SPP share the substrate preference for the type II orientation, prompted us to undertake the present study. Our results suggest strongly that S2Ps (E. coli RseP and Bacillus subtilis RasP) are involved in degradation of remnant SPs left in the bacterial cytoplasmic membrane, in contrast with the currently prevailing concept that SppA (12, 13), a protease unrelated to the S2P or the SPP family, is the enzyme responsible for signal peptide cleavage in bacteria.

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“…Like SPI, SPPA is a Ser-Lys dyad protease (76,180) that spans the membrane once, with a large C-terminal domain localized to the periplasmic space (180). However, it is different from SPI in that SPPA is inhibited by common serine protease inhibitors (63).…”

Section: Sppamentioning

confidence: 99%

Membrane Proteases in the Bacterial Protein Secretion and Quality Control Pathway

Dalbey

Wang

Dijl

2012

Microbiol Mol Biol Rev

128

129

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SUMMARY Proteolytic cleavage of proteins that are permanently or transiently associated with the cytoplasmic membrane is crucially important for a wide range of essential processes in bacteria. This applies in particular to the secretion of proteins and to membrane protein quality control. Major progress has been made in elucidating the structure-function relationships of many of the responsible membrane proteases, including signal peptidases, signal peptide hydrolases, FtsH, the rhomboid protease GlpG, and the site 1 protease DegS. These enzymes employ very different mechanisms to cleave substrates at the cytoplasmic and extracytoplasmic membrane surfaces or within the plane of the membrane. This review highlights the different ways that bacterial membrane proteases degrade their substrates, with special emphasis on catalytic mechanisms and substrate delivery to the respective active sites.

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Crystal Structure of a Bacterial Signal Peptide Peptidase

Cited by 45 publications

References 39 publications

Massive Formation of Intracellular Membrane Vesicles in Escherichia coli by a Monotopic Membrane-bound Lipid Glycosyltransferase

Massive Formation of Intracellular Membrane Vesicles in Escherichia coli by a Monotopic Membrane-bound Lipid Glycosyltransferase

Post-liberation cleavage of signal peptides is catalyzed by the site-2 protease (S2P) in bacteria

Membrane Proteases in the Bacterial Protein Secretion and Quality Control Pathway

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