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从植物细胞悬浮培养物(越橘)的一种分泌型过氧化物酶中鉴定出人类Lewis(a)碳水化合物基序。

Identification of the human Lewis(a) carbohydrate motif in a secretory peroxidase from a plant cell suspension culture (Vaccinium myrtillus L.).

作者信息

Melo N S, Nimtz M, Conradt H S, Fevereiro P S, Costa J

机构信息

ITQB/IBET, Oeiras, Portugal.

出版信息

FEBS Lett. 1997 Sep 29;415(2):186-91. doi: 10.1016/s0014-5793(97)01121-6.

Abstract

This paper reports for the first time the presence of the human Lewis(a) type determinant in glycoproteins secreted by plant cells. A single glycopeptide was identified in the tryptic hydrolysis of the peroxidase VMPxC1 from Vaccinium myrtillus L. by HPLC/ESI-MS. The oligosaccharide structures were elucidated by ESI-MS-MS and by methylation analysis before and after removal of fucose by mild acid hydrolysis. The major structure determined is of the biantennary plant complex type containing the outer chain motif Lewis(a) [structure in text]. A corresponding fucosyltransferase activity catalyzing the formation of Lewis(a) type structures in vitro was identified in cellular extracts of the suspension cultures.

摘要

本文首次报道了植物细胞分泌的糖蛋白中存在人Lewis(a)型决定簇。通过高效液相色谱/电喷雾电离质谱(HPLC/ESI-MS)在越橘(Vaccinium myrtillus L.)过氧化物酶VMPxC1的胰蛋白酶水解产物中鉴定出一种单一糖肽。通过电喷雾串联质谱(ESI-MS-MS)以及在温和酸水解去除岩藻糖前后进行甲基化分析来阐明寡糖结构。确定的主要结构是含有外链基序Lewis(a) [文中结构]的双天线植物复合类型。在悬浮培养物的细胞提取物中鉴定出一种相应的岩藻糖基转移酶活性,其在体外催化Lewis(a)型结构的形成。

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