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431.
Thirumalai R. Kannan Manickam Krishnan Kumaraguruparan Ramasamy Argentina Becker Olga N. Pakhomova P. John Hart Joel B. Baseman 《Molecular microbiology》2014,93(3):568-581
Community‐acquired respiratory distress syndrome (CARDS) toxin from Mycoplasma pneumoniae is a 591‐amino‐acid virulence factor with ADP‐ribosyltransferase (ADPRT) and vacuolating activities. It is expressed at low levels during in vitro growth and at high levels during colonization of the lung. Exposure of experimental animals to purified recombinant CARDS toxin alone is sufficient to recapitulate the cytopathology and inflammatory responses associated with M. pneumoniae infection in humans and animals. Here, by molecular modelling, serial truncations and site‐directed mutagenesis, we show that the N‐terminal region is essential for ADP‐ribosylating activity. Also, by systematic truncation and limited proteolysis experiments we identified a portion of the C‐terminal region that mediates toxin binding to mammalian cell surfaces and subsequent internalization. In addition, the C‐terminal region alone induces vacuolization in a manner similar to full‐length toxin. Together, these data suggest that CARDS toxin has a unique architecture with functionally separable N‐terminal and C‐terminal domains. 相似文献
432.
433.
A low molecular weight protein from dry mung bean (Vigna radiata) embryonic axes has been purified to near homogeneity by chromatography on DEAE-cellulose and hydroxylapatite. It shows a molecular weight of about 12,000 in sodium dodecyl sulfate-polyacrylamide gels and a sedimentation coefficient of about 2 S in sucrose gradients. This protein occurs in greater amounts in dry axes than in dry cotyledons, and it dramatically disappears during early germination of the seed. Affinity chromatography tests do not indicate it as a trypsin inhibitor or as a glycoprotein. It is a water-soluble cytoplasmic protein exhibiting an amino acid composition characteristic of storage proteins with a high content of glutamic acid/glutamine. We suggest that it is a low molecular weight storage albumin.Abbreviations Asx
aspartic acid/asparagine
- BSA
bovine serum albumin
- Con A
concanavalin A
- EB
extraction buffer
- Glx
glutamic acid/glutamine
- HA
hydroxylapatite
- PB
phosphate buffer
- SDS-PAGE
sodium dodecyl sulfate-polyacrylamide gel electrophoresis
- TCA
trichloroacetic acid 相似文献