Morris A J, Gallagher J T, Dexter T M
Cancer Research Campaign Department of Medical Oncology, Christie Hospital, Withington, Manchester, U.K.
Biomed Chromatogr. 1986 Feb;1(1):41-7. doi: 10.1002/bmc.1130010110.
The trypsin-sensitive glycopeptides from cell surfaces of a multipotential murine haemopoietic cell line (DE) have been studied using serial lectin affinity chromatography on columns of immobilized lentil lectin (LCA), concanavalin A (Con A), and wheat-germ agglutinin (WGA). WGA-binding material consisted of glycopeptides that failed to bind to LCA and Con A. Step elution from the WGA-column with 0.01-, 0.1-, 0.5- and 1.0 M N-acetyl-D-glucosamine yielded four affinity classes of glycopeptide (WGA-W, WGA-I, WGA-S and WGA-SS respectively). WGA-W, WGA-I and WGA-S contained both alkali-stable (N-linked) and alkali-labile (O-linked) carbohydrate on high molecular weight glycopeptides. The WGA-SS fraction contained only N-linked carbohydrate. N-linked glycopeptides isolated from each WGA-binding class differed in molecular size, relative N-acetylneuraminic acid content and affinity for Ricinus communis 120 agglutinin. endo-beta-Galactosidase digestion showed that these glycopeptides contained polylactosamine-type glycans. Gel filtration profiles of the enzyme treated materials were different for each WGA-binding population suggesting variation in branching patterns and/or substitution with fucose residues. Affinity chromatography has shown that the WGA binding molecules are the major glycopeptide group at DE cell surfaces.
利用固定化扁豆凝集素（LCA）、伴刀豆球蛋白A（Con A）和麦胚凝集素（WGA）柱上的连续凝集素亲和层析，对一种多能小鼠造血细胞系（DE）细胞表面的胰蛋白酶敏感糖肽进行了研究。WGA结合物质由未能与LCA和Con A结合的糖肽组成。用0.01 M、0.1 M、0.5 M和1.0 M N-乙酰-D-葡萄糖胺从WGA柱上进行分步洗脱，得到了四类亲和性糖肽（分别为WGA-W、WGA-I、WGA-S和WGA-SS）。WGA-W、WGA-I和WGA-S在高分子量糖肽上同时含有碱稳定（N-连接）和碱不稳定（O-连接）碳水化合物。WGA-SS级分仅含有N-连接碳水化合物。从每个WGA结合类别中分离出的N-连接糖肽在分子大小、相对N-乙酰神经氨酸含量以及对蓖麻凝集素120的亲和性方面存在差异。内切β-半乳糖苷酶消化表明这些糖肽含有多乳糖胺型聚糖。酶处理材料的凝胶过滤图谱对于每个WGA结合群体都不同，表明分支模式和/或岩藻糖残基取代存在差异。亲和层析表明，WGA结合分子是DE细胞表面的主要糖肽基团。
