Girardet J M, Coddeville B, Plancke Y, Strecker G, Campagna S, Spik G, Linden G
Laboratoire des BioSciences de l'Aliment, associé INRA, Université Henri Poincaré-Nancy 1, France.
Eur J Biochem. 1995 Dec 15;234(3):939-46. doi: 10.1111/j.1432-1033.1995.939_a.x.
The heat-stable acid-soluble phosphoglycoprotein component PP3 was isolated from the bovine milk proteose peptone fraction by concanavalin A affinity chromatography. Glycopeptides were released by pronase digestion of the milk component PP3 and were subsequently separated by high-pH anion-exchange chromatography on CarboPac PA-1. The primary structures of the glycan and peptide moieties of eight N-glycopeptides have been established by combining methylation analysis, mass spectrometry, 400-MHz 1H-NMR spectroscopy, and peptide sequence analysis. All the analyzed fractions contained biantennary N-acetyllactosamine-type carbohydrate chains, some of them with a GalNAc(beta 1-4)GlcNAc or a NeuAc(alpha 2-6)GalNAc(beta 1-4)GlcNAc group. This particular sequence did or did not replace the Gal(beta 1-4)GlcNAc group usually found in most N-linked glycans. Moreover, the sialylated Gal and GalNAc residues were only found on the Man(alpha 1-3) antenna.
通过伴刀豆球蛋白A亲和色谱法从牛乳蛋白酶解物组分中分离出热稳定的酸溶性磷酸糖蛋白组分PP3。通过对牛奶组分PP3进行链霉蛋白酶消化释放出糖肽，随后在CarboPac PA - 1上通过高pH阴离子交换色谱法进行分离。通过结合甲基化分析、质谱、400兆赫1H - NMR光谱和肽序列分析，确定了8种N - 糖肽的聚糖和肽部分的一级结构。所有分析的级分都含有双触角N - 乙酰乳糖胺型碳水化合物链，其中一些带有GalNAc(β1 - 4)GlcNAc或NeuAc(α2 - 6)GalNAc(β1 - 4)GlcNAc基团。这个特定序列取代或未取代大多数N - 连接聚糖中通常存在的Gal(β1 - 4)GlcNAc基团。此外，唾液酸化的Gal和GalNAc残基仅在Man(α1 - 3)触角上发现。
