Purification and refolding of the recombinant subunit B protein of the Aggregatibacter actinomycetemcomitans cytolethal distending toxin / 대한치주과학회지
The Journal of the Korean Academy of Periodontology
;
: 343-354, 2008.
Artigo
em Inglês
| WPRIM
| ID: wpr-148369
ABSTRACT
PURPOSE:
Aggregatibacter actinomycetemcomitans is associated with localized aggressive periodontitis. It produces cytolethal distending toxin (CDT), which induces cell cycle arrest in the G2/M phase. The CDT holotoxin is composed of CdtA, CdtB, and CdtC. CdtB has structural homology to human DNase I and is an active component of the CDT complex acting as a DNase. In particular, the pattern homology seen in the CdtB subunit has been associated with specific DNase I residues involved in enzyme catalysis, DNA binding, and metal ion binding. So, to study the functions and regulation of recombinant CdtB, we made up a quantity of functional recombinant CdtB and tested it in relation to the metal ion effect. MATERIALS ANDMETHODS:
We constructed the pET28a-cdtB plasmid from A. actinomycetemcomitans Y4 by genomic DNA PCR and expressed it in the BL21 (DE3) Escherichia coli system. We obtained the functional recombinant CdtB by the refolding system using the dialysis method and then analyzed the DNase activity and investigated the metal ion effect from plasmid digestion.RESULTS:
The recombinant CdtB subunit was expressed as the inclusion bodies. We were able to obtain functional recombinant CdtB subunit using refolding system. We confirmed that our refolded recombinant CdtB had DNase activity and was influenced by the metal ions Mg2+ and Ca2+.CONCLUSION:
We suggest that the factors influencing recombinant CdtB may contribute to CDT associated diseases, such as periodontitis, endocarditic, meningitis, and osteomyelitis.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Osteomielite
/
Periodontite
/
Periodontite Agressiva
/
Plasmídeos
/
Toxinas Bacterianas
/
DNA
/
Catálise
/
Corpos de Inclusão
/
Reação em Cadeia da Polimerase
/
Ácido Edético
Limite:
Humanos
Idioma:
Inglês
Revista:
The Journal of the Korean Academy of Periodontology
Ano de publicação:
2008
Tipo de documento:
Artigo
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