从猪甲状腺球蛋白中分离和分级分离糖肽

Isolation and fractionation of glycopeptides from porcine thyroglobulin.

Fukuda M, Egami F

Biochem J. 1971 Jul;123(3):407-14. doi: 10.1042/bj1230407.

Abstract

Glycopeptides were isolated by gel filtration on Sephadex G-25 and Sephadex G-50 from a Pronase digest of porcine thyroglobulin. 2. Isolated glycopeptides were separated into five main fractions on a column of DEAE-Sephadex A-25. Of these fractions I to III were further purified by SE-Sephadex C-25 or DEAE-Sephadex A-25 column chromatography. Several of the purified glycopeptides were homogeneous on paper electrophoresis. 3. Based on the chemical composition and molecular weight of the fractionated glycopeptides, two distinct types of heterosaccharide chain were demonstrated. 4. One type of the heterosaccharide unit consisted of four to eight residues of mannose and two residues of glucosamine and had a molecular weight of 1000-1700. The other type of unit contained sialic acid, fucose and galactose in addition to mannose and glucosamine and had a molecular weight of about 3600. 5. Mild alkaline treatment of the glycopeptide did not result in the destruction of threonine and serine. 2-Acetamido-1-N-(4-l-aspartyl)-2-deoxy-beta-d-glucopyranosylamine was isolated from partial acid hydrolysates.

摘要

通过在葡聚糖凝胶G - 25和葡聚糖凝胶G - 50上进行凝胶过滤，从猪甲状腺球蛋白的链霉蛋白酶消化物中分离出糖肽。2. 将分离得到的糖肽在DEAE - 葡聚糖凝胶A - 25柱上分离成五个主要部分。其中I至III部分通过SE - 葡聚糖凝胶C - 25或DEAE - 葡聚糖凝胶A - 25柱色谱进一步纯化。几种纯化后的糖肽在纸上电泳时呈均一性。3. 根据分级分离的糖肽的化学组成和分子量，证明了两种不同类型的杂糖链。4. 一种杂糖单元由四到八个甘露糖残基和两个氨基葡萄糖残基组成，分子量为1000 - 1700。另一种单元除了含有甘露糖和氨基葡萄糖外，还含有唾液酸、岩藻糖和半乳糖，分子量约为3600。5. 对糖肽进行温和的碱性处理不会导致苏氨酸和丝氨酸的破坏。从部分酸水解产物中分离出了2 - 乙酰氨基 - 1 - N - (4 - L - 天冬氨酰基) - 2 - 脱氧 - β - D - 吡喃葡萄糖胺。