Utility of pentose colorimetric assay for the purification of potato lectin, an arabinose-rich glycoprotein |
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Authors: | Siddanakoppalu N Pramod Yeldur P Venkatesh |
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Institution: | (1) Department of Biochemistry & Nutrition, Central Food Technological Research Institute (CFTRI), Mysore, 570 020, Karnataka State, India |
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Abstract: | Potato lectin (Solanum tuberosum agglutinin, STA) is an unusual glycoprotein containing approximately 50% carbohydrates by weight. Of the total carbohydrates,
92% is contributed by L-arabinose, which are O-linked to hydroxyproline residues. The ferric chloride-orcinol assay (Bial’s test), which is specific for pentoses has so
far been used only for the determination of free pentoses in biological samples. However, this colorimetric assay has not
been used for the detection of pentoses in bound form as it occurs in Solanaceae lectins (potato, tomato, and Datura lectins).
Utilizing the pentose colorimetric assay for monitoring the presence of potato lectin, a simpler and shorter procedure for
the purification of this lectin from potato tubers has been developed. The yield of potato lectin (1.73 mg per 100 g potato
tuber) is twice compared to the yields reported in earlier procedures. Although potato lectin is well known for its specificity
to free trimers and tetramers of N-acetyl-D-glucosamine (GlcNAc), it possesses a similar specificity to the core (GlcNAc)2 of N-linked glycoproteins. The utilization of the pentose assay in the purification of arabinose-rich lectins/agglutinins
obviates the necessity for the use of agglutination assay in the various purification steps. The pentose assay appears to
be a simple and convenient colorimetric assay for detecting any pentose-rich glycoprotein in plant extracts. The utility of
the pentose assay appears to have a significant potential in the detection of hydroxyproline-rich glycoproteins (HRGPs), which
are generally O-arabinosylated. |
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Keywords: | Arabinose-rich glycoprotein Ferric chloride-orcinol assay Pentose assay Potato lectin Solanum tuberosum agglutinin |
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