Chemical synthesis of globotriose and galabiose: relative stabilities of their complexes with Escherichia coli Shiga-like toxin-1 as determined by denaturation-titration with guanidinium chloride
UNSPECIFIED. (1998) Chemical synthesis of globotriose and galabiose: relative stabilities of their complexes with Escherichia coli Shiga-like toxin-1 as determined by denaturation-titration with guanidinium chloride. JOURNAL OF THE CHEMICAL SOCIETY-PERKIN TRANSACTIONS 1 (15). pp. 2287-2294. ISSN 0300-922XFull text not available from this repository.
Globotriose [alpha-D-Gal-(1-->4)-beta-D-Gal-(1-->4)-D-Glc] is the carbohydrate moiety of the globotriosyl ceramide (Gb(3)), also known as the germinal centre B-cell differentiation antigen CD77, a glycolipid present on the plasma membrane of certain mammalian cells. In Gb(3), globotriose functions as the cell-surface receptor for Shiga toxin and for the Shiga-like toxins (verocytotoxins). Here we report the chemical synthesis of globotriose and the corresponding terminal disaccharide, galabiose [alpha-D-Gal(1-->4)-beta-D-Gal]. Globotriose and galabiose are attached via a linker to CNBr-activated Sepharose to generate affinity matrices that permit the one-step purification of recombinant Shiga-like toxin-l from crude E. coli homogenates. Toxin is released from either of the immobilised saccharides by elution with 6 M guanidinium chloride. After dilution of the denaturant, the released toxin had full catalytic activity. Denaturation-titration experiments show that the bound toxin is released from galabiose-Sepharose at 2.3 M guanidinium chloride, while its release from globotriose-Sepharose requires a higher concentration of 4.8 M. These results indicate that the glucose component of globotriose contributes similar to 2.6 kcal mol(-1) to the binding;energy relative to galabiose.
|Item Type:||Journal Article|
|Subjects:||Q Science > QD Chemistry|
|Journal or Publication Title:||JOURNAL OF THE CHEMICAL SOCIETY-PERKIN TRANSACTIONS 1|
|Publisher:||ROYAL SOC CHEMISTRY|
|Official Date:||7 August 1998|
|Number of Pages:||8|
|Page Range:||pp. 2287-2294|
Actions (login required)