Wagner G F, Hampong M, Park C M, Copp D H
Gen Comp Endocrinol. 1986 Sep;63(3):481-91. doi: 10.1016/0016-6480(86)90149-8.
This report describes the isolation of teleocalcin, a glycoprotein hormone from the corpuscles of Stannius (CS) of sockeye salmon (Oncorhynchus nerka), using affinity, gel exclusion, and ion exchange chromatography. In discontinuous, gradient (10-20%) SDS gels under nonreducing conditions, teleocalcin had an estimated molecular weight of of 39,300 and migrated as a single band. Two bands with estimated molecular weights of 28,000 and 32,000 were obtained upon reduction and carboxymethylation of the molecule. In acid-urea gels, teleocalcin migrated as one band with an Rf of 0.17. Amino acid sequence analysis revealed single residues for the first 19 amino acids, with phenylalanine as the N-terminal residue. Teleocalcin did not resemble parathyroid hormone (PTH) with respect to amino acid composition and exhibited no cross-reactivity in two PTH radioimmunoassays. Gas-liquid chromatographic analysis of teleocalcin demonstrated that mannose was the principal sugar present (1.86%) and glucosamine was the only hexosamine identified (2.49%). Smaller quantities of galactose (0.49%), fucose (0.44%), and sialic acid (1.6%) were also found. In bioassays using juvenile rainbow trout (Salmo gairdneri), teleocalcin significantly reduced the rate of net branchial 45Ca uptake at dosages as low as 0.02 microgram/g body wt. This inhibitory effect was dependent upon the branchial calcium uptake cycle that has been identified in this species. Pronounced inhibition was observed during periods of high uptake whereas little or no effect was obtained during the low uptake phase. The results are discussed in relation to a possible role of teleocalcin in regulating this cycle.
本报告描述了从红大马哈鱼（Oncorhynchus nerka）斯坦尼斯小体（CS）中分离远钙素（teleocalcin）的过程，该远钙素是一种糖蛋白激素，采用了亲和色谱、凝胶排阻色谱和离子交换色谱法。在非还原条件下的不连续梯度（10 - 20%）SDS凝胶中，远钙素的估计分子量为39,300，迁移时呈单一带。对该分子进行还原和羧甲基化后，得到了两条估计分子量分别为28,000和32,000的带。在酸性尿素凝胶中，远钙素迁移为一条Rf值为0.17的带。氨基酸序列分析显示，前19个氨基酸为单残基，苯丙氨酸为N端残基。远钙素在氨基酸组成方面与甲状旁腺激素（PTH）不同，并且在两种PTH放射免疫测定中没有交叉反应性。远钙素的气液色谱分析表明，甘露糖是主要存在的糖（1.86%），葡糖胺是唯一鉴定出的己糖胺（2.49%）。还发现了较少量的半乳糖（0.49%）、岩藻糖（0.44%）和唾液酸（1.6%）。在使用幼年虹鳟（Salmo gairdneri）的生物测定中，远钙素在低至0.02微克/克体重的剂量下就能显著降低鳃对45Ca的净摄取率。这种抑制作用取决于该物种中已确定的鳃钙摄取周期。在高摄取期观察到明显的抑制作用，而在低摄取期则几乎没有或没有影响。讨论了这些结果与远钙素在调节该周期中可能作用的关系。
