S Layer Protein <i>of Clostridium tetani:</i> Purification and Properties

Takumi, Kenji; Susami, Yukie; Takeoka, Aya; Oka, Tatsuzo; Koga, Tetsuro

doi:10.1111/j.1348-0421.1991.tb01587.x

Cited by 9 publications

(12 citation statements)

References 16 publications

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“…Amino acid analysis of the two subunits showed that both the subunits lacked proline. This is a very unique feature among the amino acid compositions of other S layer proteins reported (12,20), although we (26) recently found that the S layer protein of C, tetani AO 147 was poor in Prolamine content. Whether the absence of proline is a specific character of the species or the strains is unknown at present.…”

Section: Discussionmentioning

confidence: 78%

“…The surface array on the native cell wall of C. difficile GAI 1152 was usually obscure in the negatively stained preparations because of the presence of the underlying wall peptidoglycan component (26). However, this disadvantage was overcome by subjecting the cell wall to partial autolysis (26).…”

Section: Morphological Aspects Of the S Layermentioning

confidence: 99%

“…However, this disadvantage was overcome by subjecting the cell wall to partial autolysis (26). Figure 1 shows the fine structure of the regular array on a partially autolyzed cell wall fragment with the morphological unit composed of a rhombus having each side of 7.8 nm and an interior angle of 77.8° (Fig.…”

Section: Morphological Aspects Of the S Layermentioning

confidence: 99%

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The s layer composed of two different protein subunits from Clostridium difficile GAI 1152: a simple purification method and characterization.

Hagiya

Oka

Tsuji

et al. 1992

J. Gen. Appl. Microbiol.

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Surface protein layer (S layer) of Clostridium diffccile GAI 1152 was a tetragonal array with each side of 7.8 nm and interior angle of 77.8°, which was composed of the two protein subunits with respective molecular weights of 38 kDa and 42 kDa. The S layer protein subunits were directly isolated from the whole cells by simple procedures involving extraction of the whole cells with 6 M GHCI and precipitation by 50% saturation with ammonium sulfate and purified by using DEAESepharose CL-6 B column chromatography followed by DEAE-5 PW equipped to HPLC. Amino acid analysis showed that both the subunits were acidic polypeptide and neither proline nor cysteine were present. The 38 kDa subunit exhibited a single isoelectric form (p14.0), whereas the 42 kDa subunit showed multiple isoelectric forms ranging from 5.5 to 6.3 of p1. Limited proteolysis of the both subunits with Staphylococcus aureus V8 enzyme showed quite dissimilar peptide maps between them, suggesting marked differences in primary structures.Surface protein layers (S layers) are found in various microorganisms including bacteria, rickettsia and archaeobacteria (2,9,20,21) . Since the S layers are located at the outermost surface of the cell, they are directly adjacent to the environment. The S layers, therefore, have such potential functions as demonstrated by Sleytr and Messner (20) and many other investigators (3,5,11,21,22): namely, (i) protective coat, molecular sieves, and molecular and ion traps, (ii)

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Section: Discussionmentioning

confidence: 78%

Section: Morphological Aspects Of the S Layermentioning

confidence: 99%

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The s layer composed of two different protein subunits from Clostridium difficile GAI 1152: a simple purification method and characterization.

Hagiya

Oka

Tsuji

et al. 1992

J. Gen. Appl. Microbiol.

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“…Previous work has shown that the C. tetani surface layer can be extracted by treatment of cell pellets from cultures at the mid-exponential growth phase with 4 M urea (Takumi et al, 1991). Other methods commonly used to extract SLPs include treatment with LiCl, EDTA, guanidine hydrochloride or low-pH glycine.…”

Section: Resultsmentioning

confidence: 99%

“…Extraction of surface layers was essentially performed as described (Takumi et al, 1991). Ten milliliters of cultures of C. tetani were grown overnight in anaerobic broth, diluted into 100 mL fresh broth and allowed to grow for 6 h. Cultures were centrifuged (5468 g, 15 min), resuspended in 500 mL phosphate buffered saline (PBS) and centrifuged again (17 900 g, 5 min).…”

Section: Extraction Of Surface Layersmentioning

confidence: 99%

Identification and characterization of the surface-layer protein ofClostridium tetani

Qazi

Brailsford

Wright

et al. 2007

FEMS Microbiology Letters

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Many bacterial species produce a paracrystalline layer, the surface layer, which completely surrounds the exterior of the cell. In some bacteria, the surface layer is implicated in pathogenesis. Two proteins present in cell wall extracts from Clostridium tetani have been investigated and identified one of these has been unambiguously as the surface-layer protein (SLP). The gene, slpA, has been located in the genome of C. tetani E88 that encodes the SLP. The molecular mass of the protein as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis is considerably larger than that predicted from the gene; however the protein does not appear to be glycosylated. Furthermore, analysis of five C. tetani strains, including three recent clinical isolates, shows considerable variation in the sizes of the SLP.

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S‐Layers, Microbial, Biotechnological Applications

Egelseer¹,

Ilk²,

Pum³

et al. 2009

Encyclopedia of Industrial Biotechnology

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Crystalline bacterial cell surface layers (S‐layers), a unique self‐assembly system optimized during billions of years of biological evolution, are one of the most commonly observed cell, envelope structures of prokaryotes. Although self‐assembly of molecules is an ubiquitous strategy of morphogenesis in nature, research in the area of molecular nanotechnology, nanobiotechnology, and biomimetics are only beginning to exploit its potential for the functionalization of surfaces and interfaces as well as for the production of biomimetic membranes and encapsulation systems. In this context, S‐layers fulfill key requirements for controlled assembly of supramolecular materials. As S‐layers are periodic structures, they exhibit identical physicochemical properties for each molecular unit down to the subnanometer level and possess pores of identical size and morphology. Many applications in nanobiotechnology depend on the ability of isolated native S‐layer proteins and S‐layer fusion proteins incorporating functional sequences to self‐assemble into monomolecular crystalline arrays in suspension, on a great variety of solid substrates, and on various lipid structures, including planar membranes and liposomes. S‐Layers have proven to be particularly suited as building blocks and patterning elements in a biomolecular construction kit involving all major classes of biological molecules enabling innovative approaches for the controlled ‘bottom‐up’ assembly of functional supramolecular structures and devices as required for life‐ and nonlife science applications.

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S Layer Protein of Clostridium tetani: Purification and Properties

Cited by 9 publications

References 16 publications

The s layer composed of two different protein subunits from Clostridium difficile GAI 1152: a simple purification method and characterization.

The s layer composed of two different protein subunits from Clostridium difficile GAI 1152: a simple purification method and characterization.

Identification and characterization of the surface-layer protein ofClostridium tetani

S‐Layers, Microbial, Biotechnological Applications

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