Glycosyl conjugates of biotinylated diaminopyridine applied for study of carbohydrate-to-carbohydrate interaction |
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Authors: | Natalia Utkina Seon-Joo Yoon Sen-itiroh Hakomori |
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Institution: | (1) Division of Biomembrane Research, Pacific Northwest Research Institute, 720 Broadway, Seattle, WA 98122-4302, USA;(2) Depts. of Microbiology and Global Health, University of Washington, Seattle, WA 98122, USA;(3) N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Science, Leninsky Prospect 47, Moscow, 119991, Russia;(4) Present address: Translational Research Center for Protein Function Control, Yonsei University, Seoul, 120-749, Republic of Korea; |
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Abstract: | Previous studies by us and others established that cell-cell adhesion is mediated by specific carbohydrate-to-carbohydrate
interaction (CCI). Those previous studies were based on various biochemical and biophysical approaches, including the use
of labeled glycosyl epitopes with fluorescent tag. However, these methods ideally require that the glycosyl epitope must be
fixed to a solid phase molecule, preferably with multivalency. The purpose of the present study is to establish a CCI process
using specific glycosyl residues conjugated to biotinylated diaminopyridine (BAP), and to observe: (i) clear occurrence of
homotypic CCI between “Os Fr.B” having 5–6 GlcNAc termini, vs. absence of such homotypic CCI between “Os Fr.1” having 2 GlcNAc termini; (ii) occurrence of heterotypic CCI between GM3
ganglioside and Os Fr.B, vs. absence of such heterotypic CCI between GM3 and Os Fr.1. Interaction between Os Fr.B-BAP conjugate and Os Fr.B-ceramide
mimetic (Os Fr.B-mCer) was demonstrated based on two experiments: (i) dose-dependent binding of Os Fr.B-BAP conjugate to polystyrene
plates coated with Os Fr.B-mCer was observed in the presence of bivalent cation, a prerequisite for all CCI processes, and
such binding was abolished by EDTA; (ii) binding between equal nanomolar Os Fr.B-BAP and Os Fr.B-mCer was inhibited by mM
concentration Os Fr.B without conjugate, in dose-dependent manner. Thus, cell adhesion processes based on homotypic CCI between
N-linked glycans having multiple GlcNAc termini, and heterotypic CCI between GM3 and such glycans, were clearly observed using
BAP conjugates of glycosyl epitopes. |
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