Abstract
Thirty years ago, in 1977, I unexpectedly discovered a new enzyme, glycoamidase A [1–3] that cleaves intact carbohydrate moiety from the original glycoproteins without affecting either the carbohydrate or peptide structures. This new enzyme was readily approved by the International Enzyme Committee and was assigned a new number: EC.3.5.1.52. At that time, I had been working on determining the primary amino acid sequence of stem bromelain, a pineapple proteolytic enzyme, which my professor had shown to be a glycoprotein. The presence of the sugar moieties was really an obstruction in sequencing bromelain. Therefore, I dreamed of having an enzyme that would cleave the carbohydrate chains without affecting the peptide structure. The dream came true as, all of a sudden, this enzyme appeared in front of my eyes. I was convinced that this was a gift from the Goddess of Science to a humble female scientist struggling and being frustrated in a male-dominant Japanese academic world. The new enzyme had been hiding in the crude β-glucosidase, a product supplied by Sigma Ltd. This development encouraged me to commit myself to the comprehensive analysis of N-linked oligosaccharide structures in glycoproteins. Under normal circumstances, in those days, the freedom to conduct independent research would have been difficult for a novice junior faculty member of the medical school. Fortunately, because of the sudden move of my professor out of the school, I had the opportunity to completely switch my research direction to determine the carbohydrate structures that were cleaved from several glycoproteins by the enzyme glycoamidase A. The first target was the oligosaccharide moiety of stem bromelain [4], followed by ovalbumin [5], human fibrinogen [6], proteoglycan core molecule [7], human placenta and umbilical cord [8], Taka-amylase A apoprotein [9], and xylose-containing common structural unit in N-linked oligosaccharides of laccase from sycamore cells [10].
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References
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Acknowledgments
I would like to acknowledge the work of those who directly helped to develop the N-glycan structure determination technique described here, in particular the contribution of Dr. Kay-Hooi Khoo (Taipei), Dr. Noboru Tomiya, Hiroaki Nakagawa, and Hirokazu Yagi, as well as the insightful advice of Professors Yuan-Chuan Lee (USA), Takashi Muramatsu, Yoji Arata, and Koichi Kato.
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Takahashi, N. (2011). My 30-Year Devotion to the N-Linked Oligosaccharide Structures. In: Wu, A. (eds) The Molecular Immunology of Complex Carbohydrates-3. Advances in Experimental Medicine and Biology, vol 705. Springer, Boston, MA. https://doi.org/10.1007/978-1-4419-7877-6_2
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