Determination of glycosylation sites using a protein sequencer and deglycosylation of native yeast invertase by endo-beta-N-acetylglucosaminidase. |
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Authors: | K Takegawa M Yoshikawa T Mishima M Nakoshi S Iwahara |
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Institution: | Department of Bioresource Science, Faculty of Agriculture, Kagawa University, Japan. |
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Abstract: | Endo-beta-N-acetylglucosaminidase from Arthrobacter protophormiae was tested for its capacity to release N-linked sugar chains from native yeast invertase. The enzyme liberated about 80% of the sugar chains from the native invertase. Deglycosylated invertase was digested by chymotrypsin or pepsin, and twelve N-acetylglucosamine-containing glycopeptides were isolated. The amino acid sequences of these glycopeptides were analyzed by a protein sequencer, and the elution position of 4-L-aspartylglycosylamine was directly identified by conventional sequencing. The endo-beta-N-acetylglucosaminidase was found to remove mainly nine sugar chains from native invertase. |
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