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| Shin, In-Sun; Nishikawa, Kiyotaka; Maruyama, Hiroki; Ishii, Satoshi. |
| A two-step binding assay for globotriaosylceramide (Gb3) content was developed by histidine-tagging strategy, which is a well-established method for the purification of recombinant proteins. The complete binding of the recombinant His-tagged Shiga toxin 1B subunit (1B-His) (1 microg/ml) to the standard Gb3 adsorbed on a multi-well H type plate was observed within 30 min at 37 degrees C; and its binding could be visualized by the following applications of HisProbe-HRP (8 microg/ml) and tetramethylbenzidine (TMB) peroxidase substrate. The 1B-His binding assay was linear over the range of 1 to 100 ng of Gb3 per well. The binding of 1B-His was specific to Gb3 separated from HeLa cells, and no major cross-reactivity of other glycolipids in Folch's lower... |
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Palavras-chave: Globotriaosylceramide; Shiga toxin; Binding assay method; His-tagged protein. |
| Ano: 2006 |
URL: http://ir.obihiro.ac.jp/dspace/handle/10322/833 |
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| ALVARIZ,RAFAELA M.; MOREIRA,ISABEL T.D.S.; CURY,GABRIELA K.; VARGAS,CARMEN R.; BARSCHAK,ALETHÉA G.. |
| Abstract: Fabry disease (FD) is an X-linked inherited disease and occurs due to mutations in GLA gene that encodes the α-galactosidase enzyme. Consequently, there is an accumulation of enzyme substrates, namely globotriaosylceramide (GB3). FD is a multisystemic disease, caused by storage of GB3 in vascular endothelia, with significant renal, cardiac and vascular involvement. The aim of this work was to evaluate the in vitro effect of GB3 on electron transport chain complexes (ETC) and redox parameters. Biochemical biomarkers were determined in homogenates of cerebral cortex, kidneys and liver of Wistar rats in the presence or absence of GB3 at concentrations of 3, 6, 9 and 12 mg/L. We found that GB3 caused an increase of ETC complexes II and IV activities,... |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: Electron transport chain; Fabry disease; Globotriaosylceramide; GB3; Oxidative stress; Reactive species. |
| Ano: 2019 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37652019000300710 |
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