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Plant Polysaccharide Array for Studying Carbohydrate-Binding Proteins

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Abstract

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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Abbreviations

AGA:

apiogalacturonan

CBP:

carbohydrate-binding proteins

HG:

homogalacturonan

Mw and Mn:

weight average and number average relative molecular weights, respectively

NHS:

N-hydroxysuccinimide

PBS:

phosphate buffered saline

RG-I:

rhamnogalacturonan I

RG-II:

rhamnogalacturonan II

XGA:

xylogalacturonan

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Acknowledgments

The authors are grateful to L. V. Kozlova (Kazan Institute of Biochemistry and Biophysics of Kazan Science Center of the Russian Academy of Sciences) for fruitful discussion of the results and help in preparation of the article materials and to A. F. Akhmetgalieva (Kazan Institute of Biochemistry and Biophysics of Kazan Science Center of the Russian Academy of Sciences) for technical support in preparing some commercial polysaccharides. The authors would like to express their gratitude to Dr. M.-C. Ralet and Dr. F. Gillon (French National Institute for Agricultural Research, Nantes, France) for kindly providing the sample of INRA-RU2 antibody as well as to Dr. E. A. Gunther (IPhis FRC Komi SC UB RAS) for kindly providing the polysaccharide’s samples derived from callus cultures.

Funding

This work was financially supported by the Russian Science Foundation [projects no. 20-63-47110 (printing and working with glycan arrays) and no. 20-64-47036 (purification, separation and characterization of polysaccharides)], as well as by the State Assignment no. AAAA-A18-118022790083-9 (selection of a base of commercial polysaccharides).

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Authors and Affiliations

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

    Anna V. Nikiforova, Nikolai V. Bovin & Nadezhda V. Shilova

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

    Victoria V. Golovchenko & Olga A. Patova

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

    Polina V. Mikshina & Tatyana A. Gorshkova

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  1. Anna V. Nikiforova
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  2. Victoria V. Golovchenko
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  3. Polina V. Mikshina
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  4. Olga A. Patova
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  5. Tatyana A. Gorshkova
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  6. Nikolai V. Bovin
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  7. Nadezhda V. Shilova
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Contributions

T. A. Gorshkova and N. V. Bovin: problem statement and supervision; N. V. Shilova: development of the concept of the article, formation and discussion of the research results; A. N. Nikiforova: carrying out experiments, description and graphical representation of results, formatting the article; V. V. Golovchenko, O. A. Patova, and P. V. Mikshina: obtaining, purification, fractionation, and characterization of polysaccharide samples, formatting the results in this part of the study, discussion and text editing.

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Correspondence to Anna V. Nikiforova.

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The authors declare no conflicts of interest in financial or any other sphere. This article does not contain any studies with human participants or animals performed by any of the authors.

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Nikiforova, A.V., Golovchenko, V.V., Mikshina, P.V. et al. Plant Polysaccharide Array for Studying Carbohydrate-Binding Proteins. Biochemistry Moscow 87, 890–902 (2022). https://doi.org/10.1134/S0006297922090036

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