PURIFICATION AND CHARACTERIZATION OF A SULFATED GLYCOPROTEIN SECRETED BY SERTOLI CELLS
PURIFICATION AND CHARACTERIZATION OF A SULFATED GLYCOPROTEIN SECRETED BY SERTOLI CELLS
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DOI：
10.1021/bi00371a003
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发表时间：
1986-11-18
期刊：
影响因子：
2.9
通讯作者：
BISHOP, P
中科院分区：
文献类型：
--
作者：
GRISWOLD, MD;ROBERTS, K;BISHOP, P
硫酸化糖蛋白2（SGP-2），支持细胞的主要分泌产物，从细胞培养基中纯化，通过反相高效液相色谱法。天然蛋白质由41000和29000道尔顿的二硫键连接的单体组成，这些单体具有强烈的结合成多聚体的倾向。对纯化的SGP-2进行氨基酸分析，并含有高水平的Asx（11.1%）、Glx（15.1%）和亮氨酸（11.5%）。对纯化的SGP-2上的寡糖进行分析以确定单糖组成和完整碳水化合物部分的分子量。SGP-2显示为23.7%碳水化合物，并且由1%岩藻糖、3.5%甘露糖、4.1%半乳糖、7.1% N-乙酰葡糖胺和8.0% N-乙酰神经氨酸组成。未检出N-乙酰半乳糖胺。当用蛋白酶消化SGP-2时，在Bio-Gel P-6柱上对完整的寡糖进行色谱分析，发现在约3300 g/mol的单个对称峰中沉淀。根据这些结果，SGP-2上的寡糖被认为是由类似于胎球蛋白上发现的三触角链组成。用链霉蛋白酶消化~（35）SO_（42）-标记的SGP-2，可通过层析从寡糖中分离出游离氨基酸。35 SO 42-被证明与SGP-2的寡糖部分相关。
Sulfated glycoprotein 2 (SGP-2), the major secretion product of Sertoli cells, was purified from cell culture medium by reverse-phase high-performance liquid chromatography. The native protein consists of disulfide-linked monomers of 41000 and 29000 daltons which have a strong tendency to associate into multimers. The purified SGP-2 was subjected to amino acid analysis and contained high levels of Asx (11.1%), Glx (15.1%), and leucine (11.5%). The oligosaccharides on the purified SGP-2 were analyzed to determined the monosaccharide compositions and the molecular weights of the intact carbohydrate moieties. SGP-2 was shown to be 23.7% carbohydrate and consisted of 1% fucose, 3.5% mannose, 4.1% galactose, 7.1% N-acetylglucosamine, and 8.0% N-acetylneuraminic acid. No N-acetylgalactosamine was detected. When the SGP-2 was digested with proteases, the intact oligosaccharides were chromatographed over a Bio-Gel P-6 column and found to elute in a single symmetrical peak of approximately 3300 g/mol. On the basis of these results, the oligosaccharides on SGP-2 were proposed to consist of triantennary chains similar to those found on fetuin. When the 35SO42--labeled SGP-2 was digested with Pronase, the free amino acids could be separated by chromatography from the oligosaccharide. The 35SO42- was shown to be associated with the oligosaccharide portion of SGP-2.