Crystalline bacterial cell surface layers: general principles and application potential

Sleytr, Uwe B.; Messner, Paul; Pum, Dietmar; Sára, Margit

doi:10.1111/j.1365-2672.1993.tb04339.x

Cited by 30 publications

(13 citation statements)

References 69 publications

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“…In the case of ORF1p, proline is the most abundant residue, and the central domain has a quite atypical, very hydrophilic composition, which probably affects binding of SDS and electro- phoretic migration. Further deviation from regular electrophoretic behavior may be expected if ORF1p is glycosylated, as are many bacterial cell surface proteins (35). No change in electrophoretic migration was observed upon heating the sample in 2% SDS ϩ 10 mM EDTA prior to electrophoresis (data not shown).…”

Section: Resultsmentioning

confidence: 94%

OlpB, a new outer layer protein of Clostridium thermocellum, and binding of its S-layer-like domains to components of the cell envelope

et al. 1995

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Several proteins of Clostridium thermocellum possess a C-terminal triplicated sequence related to bacterial cell surface proteins. This sequence was named the SLH domain (for S-layer homology), and it was proposed that it might serve to anchor proteins to the cell surface (A. Lupas, H. Engelhardt, J. Peters, U. Santarius, S. Volker, and W. Baumeister, J. Bacteriol. 176:1224-1233, 1994). This hypothesis was investigated by using the SLH-containing protein ORF1p from C. thermocellum as a model. Subcellular fractionation, immunoblotting, and electron microscopy of immunocytochemically labeled cells indicated that ORF1p was located on the surface of C. thermocellum. To detect C. thermocellum components interacting with the SLH domains of ORF1p, a probe was constructed by grafting these domains on the C terminus of the MalE protein of Escherichia coli. The SLH domains conferred on the chimeric protein (MalE-ORF1p-C) the ability to bind noncovalently to the peptidoglycan of C. thermocellum. In addition, 125I-labeled MalE-ORF1p-C was shown to bind to SLH-bearing proteins transferred onto nitrocellulose, and to a 26-to 28-kDa component of the cell envelope. These results agree with the hypothesis that SLH domains contribute to the binding of exocellular proteins to the cell surface of bacteria. The gene carrying ORF1 and its product, ORF1p, are renamed olpB and OlpB (for outer layer protein B), respectively.

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Section: Resultsmentioning

confidence: 94%

OlpB, a new outer layer protein of Clostridium thermocellum, and binding of its S-layer-like domains to components of the cell envelope

et al. 1995

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“…Although S-layers were discovered over 40 years ago, the current state of knowledge concerning their structure is somewhat primitive (6,55). So far, the highest spatial resolution models (ca.…”

Section: Discussionmentioning

confidence: 99%

Linker mutagenesis of the Caulobacter crescentus S-layer protein: toward a definition of an N-terminal anchoring region and a C-terminal secretion signal and the potential for heterologous protein secretion

1997

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Linker insertion mutagenesis was used to modify the paracrystalline surface layer (S-layer) protein (RsaA) of the gram-negative bacterium Caulobacter crescentus. Eleven unique BamHI linker insertions in the cloned rsaA gene were identified; at the protein level, these linker insertions introduced 4 to 6 amino acids at positions ranging from the extreme N terminus to the extreme C terminus of the 1,026-amino-acid RsaA protein. All linker-peptide insertions in the RsaA N terminus caused the secreted protein to be shed into the growth medium, suggesting that the RsaA N terminus is involved in cell surface anchoring. One linker-peptide insertion in the RsaA C terminus (amino acid 784) had no effect on S-layer biogenesis, while another (amino acid 907) disrupted secretion of the protein, suggesting that RsaA possesses a secretion signal lying C terminal to amino acid 784, near or including amino acid 907. Unlike extreme N-or C-terminal linker-peptide insertions, those more centrally located in the RsaA primary sequence had no apparent effect on S-layer biogenesis. By using a newly introduced linker-encoded restriction site, a 3 fragment of the rsaA gene encoding the last 242 C-terminal amino acids of the S-layer protein was expressed in C. crescentus from heterologous Escherichia coli lacZ transcription and translation initiation information. This C-terminal portion of RsaA was secreted into the growth medium, confirming the presence of a C-terminal secretion signal. The use of the RsaA C terminus for the secretion of heterologous proteins in C. crescentus was explored by fusing 109 amino acids of an envelope glycoprotein from infectious hematopoietic necrosis virus, a pathogen of salmonid fish, to the last 242 amino acids of the RsaA C terminus. The resulting hybrid protein was successfully secreted into the growth medium and accounted for 10% of total protein in a stationary-phase culture. Based on these results and features of the RsaA primary sequence, we propose that the C. crescentus S-layer protein is secreted by a type I secretion system, relying on a stable C-terminal secretion signal in a manner analogous to E. coli ␣-hemolysin, the first example of an S-layer protein secreted by such a pathway.The outer membrane of the dimorphic gram-negative bacterium Caulobacter crescentus is covered by a protein surface layer (S-layer) composed of a single 1,026-amino-acid protein termed RsaA (13). The S-layer is paracrystalline in nature, exhibiting an array of ring-like subunits (each composed of six copies of RsaA) arranged on a lattice with p6 symmetry and interlinked at the threefold rotational axis (57); proper crystallization of the S-layer is dependent on Ca 2ϩ ions (62, 63). The association between RsaA and the outer membrane is not completely understood, but the protein appears to be anchored to the outer membrane via noncovalent interactions with a specific smooth lipopolysaccharide (LPS) molecule (62). So far, the only known function of the C. crescentus S-layer is to protect cells against predation by a Bde...

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“…The considerable body of knowledge accumulated on structure, chemistry, morphogenesis, genetics and physicochemical surface properties of S‐layers has led to a variety of applications during the last decade which are summed up in Table 3 and reviewed in references [14, 48–55].…”

Section: Application Potentialmentioning

confidence: 99%

I. Basic and applied S-layer research: an overview

Sleytr¹

1997

FEMS Microbiol Rev

107

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S‐layers are crystalline monomolecular assemblies of protein or glycoprotein, which represent one of the most common cell surface structures in Archaea and Bacteria. As porous lattices completely covering the cell surface they can provide prokaryotic cells with selection advantages by functioning as protective coats, as structures involved in cell adhesion and surface recognition, as molecule or ion traps, and molecular sieves. In Archaea, which possess S‐layers as exclusive cell wall component, the (glyco)protein lattices function as cell shape determining/maintaining framework. Studies on structure, chemistry, genetics, assembly and function of S‐layers revealed a considerable application potential for the regular arrays in biotechnology, biomimetics, biomedicine and molecular nanotechnology.

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Crystalline bacterial cell surface layers: general principles and application potential

Cited by 30 publications

References 69 publications

OlpB, a new outer layer protein of Clostridium thermocellum, and binding of its S-layer-like domains to components of the cell envelope

OlpB, a new outer layer protein of Clostridium thermocellum, and binding of its S-layer-like domains to components of the cell envelope

Linker mutagenesis of the Caulobacter crescentus S-layer protein: toward a definition of an N-terminal anchoring region and a C-terminal secretion signal and the potential for heterologous protein secretion

I. Basic and applied S-layer research: an overview

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