Plant Polysaccharide Array for Studying Carbohydrate-Binding Proteins

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Anna V. Nikiforova^1,a*, Victoria V. Golovchenko², Polina V. Mikshina³, Olga A. Patova², Tatyana A. Gorshkova³, Nikolai V. Bovin¹, and Nadezhda V. Shilova¹

¹Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences, 117997 Moscow, Russia

²Institute of Physiology of Komi Science Centre of the Ural Branch of the Russian Academy of Sciences, FRC Komi SC UB RAS, 167982 Syktyvkar, Russia

³Kazan Institute of Biochemistry and Biophysics of FRC Kazan Scientific Center of the Russian Academy of Sciences, 420111 Kazan, Russia

^* To whom correspondence should be addressed.

Received June 2, 2022; Revised June 20, 2022; Accepted June 20, 2022

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The specificity of the most plant carbohydrate-binding proteins (CBP), many of which are known only through bioinformatic analysis of the genome, has either not been studied at all or characterized to a limited extent. The task of deciphering the carbohydrate specificity of the proteins can be solved using glycoarrays composed of many tens or even hundreds of glycans immobilized on a glass surface. Plant carbohydrates are the most significant natural ligands for plant proteins; this work shows that plant polysaccharides without additional modification can be immobilized on the surface, bearing N-hydroxysuccinimide activated carboxyl groups. As a result, an array of 113 well-characterized polysaccharides isolated from various plant cell walls, 23 mono- and oligosaccharides – components of polysaccharides, and glycans – ligands for widely known plant lectins was designed. Upon chemical immobilization of polysaccharides, their functional activity was preserved, which was confirmed by the results of interaction with antibodies and the plant lectin ricin. Using the constructed array, a previously unknown ability of ricin to bind polysaccharides was found, which significantly expands the knowledge of its specificity, and it was also found that a large variety of antibodies to plant polysaccharides are present in human peripheral blood.

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KEY WORDS: carbohydrate-binding proteins, plant lectins, plant polysaccharides, pectins, polysaccharide chip, glycoarray, INRA-RU2, anti-glycan human antibodies

DOI: 10.1134/S0006297922090036

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