识别和描述人类血清clusterin糖基化的网站。
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卡普隆JT, Hilliard通用,Lakins约,Tenniswood MP,西卡,卡尔SA Crabb JW
识别和描述人类血清clusterin糖基化的网站。
蛋白质科学。1997年10月,6(10):2120 - 33所示。
- PubMed ID
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9336835 (在PubMed]
- 文摘
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Clusterin是无处不在的,heterodimeric糖蛋白与多个可能的功能可能受到糖基化的影响。识别低聚糖附件网站和低聚糖的结构表征人类血清clusterin一直由质谱和埃德曼降解。Matrix-assisted激光解吸电离质谱分析显示两种分子量的holoclusterin(58505 + / - 250和63507 + / - 200)。质谱分析还揭示了分子异质性与凝聚素的α和β亚基与存在多个改变一致。数据表明,clusterin包含17 - 27%碳水化合物的重量,α亚基包含0 - 30%碳水化合物和碳水化合物β亚基包含27 - 30%。液相色谱电喷雾质谱分析了碰撞能量扫描用于选择性地识别并制备分离胰蛋白酶的糖肤。埃德曼序列分析被用来证实糖肤的身份和定义附件网站在每个肽。总共六个N-linked糖基化网站,三个α亚基的α(阿尔法64 n,阿尔法81 n, 123 n)和三个β亚基(ββ64 n,β127 n, 147 n)。七种不同的可能的低聚糖结构类型被确定质量包括:monosialobiantennary结构,bisialobiantennary结构没有或海藻糖,trisialotriantennary结构没有或海藻糖,并可能trisialotriantennary结构有两个岩藻糖和/或tetrasialotriantennary结构。网站β64 n至少表现出糖基化多样性,发现有两个类型的低聚糖、β和站点147 n表现出最大的多样性,与五、六发现类型的低聚糖。 Overall, the most abundant glycoforms detected were bisialobiantennary without fucose and the least abundant were monosialobiantennary, trisialotriantennary with two fucose and/or tetrasialotriantennary. Clusterin peptides accounting for 99% of the primary structure were identified from analysis of the isolated alpha and beta subunits, including all Ser- and Thr-containing peptides. No evidence was found for the presence of O-linked or sulfated oligosaccharides. The results provide a molecular basis for developing a better understanding of clusterin structure-function relationships and the role clusterin glycosylation plays in physiological function.