Identification of disulfide bonds and site-specific glycosylation in chicken alpha1-acid glycoprotein by matrix-assisted laser desorption ionization time-of-flight mass spectrometry.
Identification of disulfide bonds and site-specific glycosylation in chicken alpha1-acid glycoprotein by matrix-assisted laser desorption ionization time-of-flight mass spectrometry.
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DOI：
10.1016/j.ab.2004.04.041
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发表时间：
2004-08
影响因子：
2.9
通讯作者：
H. Matsunaga;Y. Sadakane;J. Haginaka
中科院分区：
文献类型：
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作者：
H. Matsunaga;Y. Sadakane;J. Haginaka
最近，我们报道了鸡α1-酸性糖蛋白的氨基酸序列。生物群落。[中英文摘要]Re.Commun.295(2002)587]。在本研究中，我们利用氨甲基化鸡肉α1-琼脂糖蛋白、氨基甲基化完全脱糖鸡肉α1-琼脂糖蛋白(CD-α1-琼脂糖基化蛋白)和非还原变性CD-α1-琼脂糖基化蛋白的胰酶切结合基质辅助激光解吸电离飞行时间质谱仪定位了鸡肉α1-琼脂糖蛋白中的二硫键和位点特异性糖基化。根据对m/z 3037.4(氨基酸序列69-76+161-183)和3453.3(氨基酸序列69-80+161-183)的检测，鸡α1-AGP的两个二硫键分别位于Cys 6-Cys 146和Cys 73-Cys 163。结果还表明ASN 16、70、77和87是完全糖基化的，ASN 62是部分糖基化的。
Recently, we reported the amino acid sequence of chicken α1-acid glycoprotein (chicken α1-AGP) [Biochem. Biophys. Res. Commun. 295 (2002) 587]. In this study, we located the disulfide bonds and site-specific glycosylation in chicken α1-AGP using tryptic digests of carbamidomethylated chicken α1-AGP, carbamidomethylated completely deglycosylated chicken α1-AGP (cd-α1-AGP), and nonreduced denatured cd-α1-AGP by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Based on the detection of peptides m/z 3037.4 (amino acid sequences 69–76 plus 161–183) and 3453.3 (amino acid sequences 69–80 plus 161–183), the two disulfide bonds of chicken α1-AGP were determined to be located at Cys 6–Cys 146 and Cys 73–Cys 163. The results also showed that Asn 16, 70, 77, and 87 were fully glycosylated and that Asn 62 was partially glycosylated.