Demonstration and preliminary characterization of a regular array in the cell wall of Clostridium difficile

Kawata, Tomio

doi:10.1016/0378-1097(84)90273-8

Cited by 22 publications

(41 citation statements)

References 8 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Exterior to a typical Gram-positive cell wall is the C. difficile S-layer, a paracrystalline proteinaceous array that surrounds the bacterium (7). The S-layer is composed of two proteins, the high molecular weight (HMW) 2 and low molecular weight (LMW) S-layer proteins (SLPs), derived by post-translational extracellular cleavage of the precursor SlpA (8).…”

mentioning

confidence: 99%

Clostridium difficile Has Two Parallel and Essential Sec Secretion Systems

Fagan

Fairweather

2011

Journal of Biological Chemistry

222

316

View full text Add to dashboard Cite

Protein translocation across the cytoplasmic membrane is an essential process in all bacteria. The Sec system, comprising at its core an ATPase, SecA, and a membrane channel, SecYEG, is responsible for the majority of this protein transport. Recently, a second parallel Sec system has been described in a number of Gram-positive species. This accessory Sec system is characterized by the presence of a second copy of the energizing ATPase, SecA2; where it has been studied, SecA2 is responsible for the translocation of a subset of Sec substrates. In common with many pathogenic Gram-positive species, Clostridium difficile possesses two copies of SecA. Here, we describe the first characterization of the C. difficile accessory Sec system and the identification of its major substrates. Using inducible antisense RNA expression and dominant-negative alleles of secA1 and secA2, we demonstrate that export of the S-layer proteins (SLPs) and an additional cell wall protein (CwpV) is dependent on SecA2. Accumulation of the cytoplasmic precursor of the SLPs SlpA and other cell wall proteins was observed in cells expressing dominant-negative secA1 or secA2 alleles, concomitant with a decrease in the levels of mature SLPs in the cell wall. Furthermore, expression of either dominant-negative allele or antisense RNA knockdown of SecA1 or SecA2 dramatically impaired growth, indicating that both Sec systems are essential in C. difficile.

show abstract

mentioning

confidence: 99%

Clostridium difficile Has Two Parallel and Essential Sec Secretion Systems

Fagan

Fairweather

2011

Journal of Biological Chemistry

222

316

View full text Add to dashboard Cite

show abstract

“…We demonstrated the presence of a squarely arranged regular array (RA) in the outer wall layer of C. difficile GAI0714 composed of two major proteins with molecular weights of 32,000 (32K) and 45,000 (45K) (11). Similar RAs were also observed on the cell walls of the other 9 strains of the organism including ATCC 11011.…”

mentioning

confidence: 61%

Purification and Immunochemical Properties of a Wall Protein Antigen from Clostridium difficile ATCC 11011

Takumi

Takeoka

Kawata

1987

Microbiology and Immunology

View full text Add to dashboard Cite

A wall-surface protein antigen, designated 32K antigen, was extracted from whole cells of Clostridium difficile strain ATCC 11011 with phosphate buffered saline and purified by ion-exchange chromatography, gel filtration, and chromatofocusing. The 32K antigen preparation was determined to be highly homogeneous by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The amino acid composition of the antigen was characteristic in the predominance of the acidic amino acids, the very low contents of methionine and histidine, and the lack of cysteine.

show abstract

“…Effect of Various Agents on Removal of the Major Cell Wall Proteins C. difficile GAI 4131 possesses 38 kDa and 42 kDa major cell wall proteins as described previously (13). The isolated cell walls were treated with hydrophobic bond-disrupting agents, chelating agents or NaCl, and subjected to SDS-PAGE to examine their effects on removal of the major proteins (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Subunit proteins isolated from some of the regular arrays have the ability to reassemble into lattices having the same dimensions as those observed on intact cells (25). Usually, the subunit proteins isolated from gram-positive bacteria require neither specific cations nor supporting layers for their reassembly, while those from gram-negative bacteria require both cations and supporting layers of Clostridium difficile (13). We also found that these strains carrying regular arrays possessed two major cell wall proteins (13), and one of the major proteins with a molecular weight of 32 kDa of strain ATCC 11011 of the organism was purified and characterized (27).…”

mentioning

confidence: 97%

“…Usually, the subunit proteins isolated from gram-positive bacteria require neither specific cations nor supporting layers for their reassembly, while those from gram-negative bacteria require both cations and supporting layers of Clostridium difficile (13). We also found that these strains carrying regular arrays possessed two major cell wall proteins (13), and one of the major proteins with a molecular weight of 32 kDa of strain ATCC 11011 of the organism was purified and characterized (27). This paper describes properties of the two major cell wall proteins with molecular weights of 38 kDa and 42 kDa, and reassembly of a tetragonal array of C. difficile GAI 4131.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Characterization and Reassembly of a Regular Array in the Cell Wall of Clostridium difficile GAI 4131

Masuda

Itoh

Kawata

1989

Microbiology and Immunology

View full text Add to dashboard Cite

The cell wall of Clostridium difficile GAI 4131 was revealed by electron microscopy to have an outer layer composed of a nearly square array and contained the two major proteins with molecular weights of 38 kDa and 42 kDa. The properties and reassembly of the two major proteins into the regular array were investigated. When the isolated cell walls were treated with hydrophobic bond-disrupting agents or a chelating agent specific for Ca2+, the two major proteins were effectively removed and the regularly arranged outer layer disappeared. The amino acid composition of the two major proteins differed from each other. The two major proteins also gave different peptide maps from each other upon proteolysis with Staphylococcus aureus V8 protease. The major proteins solubilized from the isolated cell walls with 8 M urea or 4 M guanidine hydrochloride could be reassembled into open-ended cylinders possessing the native regular pattern by dialysis against neutral buffer containing 5 mm CaCl2. The reassembled cylinders purified by centrifugation on a Percoll density gradient were composed of almost equal amounts of the 38 kDa and 42 kDa proteins ond freed from the other proteins. These results suggest that the regular array in the outer cell wall layer is constructed from the two major cell wall proteins and requires Ca2+ for its assembly.

show abstract

Demonstration and preliminary characterization of a regular array in the cell wall of Clostridium difficile

Cited by 22 publications

References 8 publications

Clostridium difficile Has Two Parallel and Essential Sec Secretion Systems

Clostridium difficile Has Two Parallel and Essential Sec Secretion Systems

Purification and Immunochemical Properties of a Wall Protein Antigen from Clostridium difficile ATCC 11011

Characterization and Reassembly of a Regular Array in the Cell Wall of Clostridium difficile GAI 4131

Contact Info

Product

Resources

About