Subunit-specific sulphation of oligosaccharides relating to charge-heterogeneity in porcine lutrophin isoforms.
Subunit-specific sulphation of oligosaccharides relating to charge-heterogeneity in porcine lutrophin isoforms.
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DOI：
10.1093/glycob/2.3.225
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发表时间：
1992-06
期刊：
影响因子：
4.3
通讯作者：
M. Ujihara;K. Yamamoto;K. Nomura;S. Toyoshima;H. Demura;Y. Nakamura;K. Ohmura;T. Osawa
中科院分区：
文献类型：
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作者：
M. Ujihara;K. Yamamoto;K. Nomura;S. Toyoshima;H. Demura;Y. Nakamura;K. Ohmura;T. Osawa
促黄体素 (LH) 由一系列具有不同电荷和生物活性的亚型组成。这项研究的目的是明确阐明 α 和 β 亚基寡糖如何促进 LH 同工型电荷。通过等电聚焦 (IEF) 将猪 LH (pLH) 分为四种亚型，然后进行亚基分离。它们的寡糖通过肼解作用释放，用 NaB3H4 还原进行标记，并通过 HPLC 使用 Mono Q 柱分级为硫酸盐 (S) 和唾液酸 (N) 残基数量不同的五个群体，指定为中性、N-1、S-1、S-N 和 S-2。低聚糖主要是硫酸化的（S-1 和 S-2），很少是唾液酸化的（N-1 和 S-N）。进一步的分析，包括刀豆球蛋白 A (Con A) 亲和层析、脱唾液酸化、脱硫、连续外切糖苷酶消化和甲基化，阐明了酸性寡糖的结构。全部都是双触角复合体类型。它们的两个外周分支是S-1中的SO4-4GalNAc beta 1-4Glc-NAc和GalNAc beta 1-4GlcNAc或GlcNAc，S-N中的SO4-4GalNAc beta 1-4GlcNAc和Sia alpha 2-6Gal beta 1-4GlcNAc，以及S-2中的(SO4-4GalNAc beta 1-4GlcNAc)2（其中GalNAc 是 N-乙酰半乳糖胺，GlcNAc 是 N-乙酰葡糖胺）。百分之十的 S-1 和 S-N 具有平分 GlcNAc 残基。两个亚基的硫酸盐残留量几乎相同；然而，α和β亚基的硫酸化方式不同。 S-1 在 α 亚基中占主导地位，而 S-1 和 S-2 是 β 亚基中的主要成分。（摘要截断为 250 字）
Lutrophin (LH) consists of an array of isoforms with different charges and bioactivities. This study was undertaken to clarify specifically how oligosaccharides of alpha and beta subunits contribute to LH isoform charges. Porcine LH (pLH) was separated into four isoforms by isoelectric focusing (IEF), followed by subunit isolation. Their oligosaccharides were released by hydrazinolysis, labelled by reduction with NaB3H4, and fractionated by HPLC with a Mono Q column into five populations differing in the number of sulphate (S) and sialic acid (N) residues, designated as Neutral, N-1, S-1, S-N and S-2. Oligosaccharides were predominantly sulphated (S-1 and S-2) and infrequently sialylated (N-1 and S-N). Further analysis, including concanavalin A (Con A) affinity chromatography, desialylation, desulphation, sequential exoglycosidase digestion and methylation, clarified the structures of the acidic oligosaccharides. All were of the biantennary complex type. Their two peripheral branches were SO4-4GalNAc beta 1-4Glc-NAc and GalNAc beta 1-4GlcNAc or GlcNAc in S-1, SO4-4GalNAc beta 1-4GlcNAc and Sia alpha 2-6Gal beta 1-4GlcNAc in S-N, and (SO4-4GalNAc beta 1-4GlcNAc)2 in S-2 (where GalNAc is N-acetylgalactosamine and GlcNAc is N-acetylglucosamine). Ten percent of S-1 and of S-N had a bisecting GlcNAc residue. Sulphate residues occurred in nearly the same amount for both subunits; however, the alpha and beta subunits were sulphated differently. S-1 predominated in the alpha subunit, while S-1 and S-2 were major components in the beta subunit.(ABSTRACT TRUNCATED AT 250 WORDS)