Affinities of Shiga toxins 1 and 2 for univalent and oligovalent Pk-trisaccharide analogs measured by electrospray ionization mass spectrometry

被引:35
作者
Kitova, Elena N.
Kitov, Pavel I.
PaszkiewiCZ, Eugenia
Kim, Jonghwa
Mulvey, George L.
Armstrong, Glen D.
Bundle, David R.
Klassen, John S. [1 ]
机构
[1] Univ Alberta, Dept Chem, Edmonton, AB T6G 2G2, Canada
[2] Univ Calgary, Fac Med, Dept Microbiol & Infect Dis, Calgary, AB T2N 4N1, Canada
基金
加拿大自然科学与工程研究理事会;
关键词
association constants; electrospray ionization mass; spectrometry; protein-oligosaccharide complexes; Shiga toxins; stoichiometry;
D O I
10.1093/glycob/cwm081
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The binding stoichiometry and affinities of the Shiga toxins, Stx1 and Stx2, for a series of uni- and oligovalent analogs of the Pk-trisaccharide were measured using the direct electrospray ionization mass spectrometry (ES-MS) assay. Importantly, it is shown that, for a given ligand, Stx1 and Stx2 exhibit similar affinities. The binding data suggest a high degree of similarity in the spatial arrangement and structural characteristics of the Pk binding sites in Stx1 and Stx2. The results confirm that both toxins recognize the alpha-D-Galp(1 -> 4)-beta-D-Galp(1 -> 4)-beta-D-Glcp carbohydrate motif of the cell surface glycolipid Gb3. This, taken together with the results of the chemical mapping study, suggests that the nature of the Pk binding interactions with Stx1 and Stx2 are similar. The affinities of Stx1-B-5 and Stx2 for the multivalent ligands reveals that site 2 of Stx2, which shares the same spatial arrangement as site 2 in Stx1, is the primary Pk binding site and that site 1 of Stx1 and of Stx2 can also participate in Pk binding.
引用
收藏
页码:1127 / 1137
页数:11
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