Purification, characterization, and bioassay of prolactin and growth hormone from temperate basses, genus Morone.
Purification, characterization, and bioassay of prolactin and growth hormone from temperate basses, genus Morone.
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温带鲈鱼属莫罗尼催乳素和生长激素的纯化、表征和生物测定。
DOI：
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发表时间：
2000
影响因子：
2.7
通讯作者：
Craig V. Sullivan
中科院分区：
文献类型：
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作者：
Leslie F. Jackson;Penny Swanson;Cunming Duan;S. Fruchtman;Craig V. Sullivan
从条纹鲈鱼（Morone saxatilis）及其与白色鲈鱼（M. saxatilis x M.通过在碱性条件下的凝胶过滤色谱法，然后通过反相高压液相色谱法，在物种之间显示相似。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳后，纯化GH的次要形式（GH I）和主要形式（GH II）均显示为单一条带（M（r）约23，000），纯化PRL也是如此（M（r）约24，000）。通过MALDI TOF质谱法测定的GH II和PRL的分子量分别为21.2和21.3 kDa。在Western印迹实验中，抗罗非鱼（Oreochromis mossambicus）24 K PRL的抗血清特异性地识别Morone PRL，而抗罗非鱼GH的抗血清特异性地识别Morone GH I和II。通过N-末端肽测序进一步证实了推定的PRL和GH I的化学身份，而对GH II进行了内部序列分析，因为它在其N-末端被阻断。经过29个氨基酸的分析，发现Morone PRL与罗非鱼24 K PRL有76%相同，与罗非鱼20 K PRL有72%相同，与鲑鱼（Oncorhynchus凯塔）PRL I有72%相同，与鳗鱼（Anguilla japonica）PRL I有69%相同。杂交条纹鲈鱼GH序列与其他几种高级海洋硬骨鱼GH序列的比对表明GH I（40个氨基酸）的序列同一性为75-85%，GH II（45个氨基酸）的序列同一性为95-98%。条纹鲈鱼生长激素II的生物活性得到证实，使用异源在体外测定的胰岛素样生长因子I mRNA的生产银鲑（On. kisutch）肝细胞。在体内的生物测定，包括垂体切除的混合条纹鲈鱼和治疗的鱼保持在淡水中与同源PRL，证实了纯化的条纹鲈鱼PRL也具有生物活性。
Prolactin (PRL) and two variants of growth hormone (GH), purified from pituitaries of striped bass (Morone saxatilis) and its hybrid with white bass (M. saxatilis x M. chrysops) by gel filtration chromatography under alkaline conditions followed by reversed-phase high pressure liquid chromatography, appear similar between species. Both the minor (GH I) and the major (GH II) forms of purified GH appeared as single bands (M(r) approximately 23,000) after sodium dodecyl sulfate-polyacrylamide gel electrophoresis, as did the purified PRL (M(r) approximately 24,000). The molecular weights of GH II and PRL determined by MALDI TOF mass spectroscopy were 21.2 and 21.3 kDa, respectively. In Western blotting experiments, an antiserum against tilapia (Oreochromis mossambicus) 24K PRL specifically recognized Morone PRL, while an antiserum against tilapia GH specifically recognized Morone GH I and II. Chemical identities of the putative PRL and GH I were further confirmed by N-terminal peptide sequencing, while internal sequence analysis was performed on GH II because it was blocked at its N-terminus. Over a stretch of 29 amino acids, Morone PRL was found to be 76% identical to tilapia 24K PRL, 72% identical to tilapia 20K PRL, 72% identical to chum salmon (Oncorhynchus keta) PRL I, and 69% identical to eel (Anguilla japonica) PRL I. Alignment of the hybrid striped bass GH sequences with those of several other advanced marine teleosts indicated 75-85% sequence identity for GH I (40 amino acids) and 95-98% identity for GH II (45 amino acids). Biological activity of striped bass GH II was confirmed using a heterologous in vitro assay of insulin-like growth factor I mRNA production by coho salmon (On. kisutch) hepatocytes. An in vivo bioassay, involving hypophysectomy of hybrid striped bass and treatment of the fish maintained in fresh water with homologous PRL, confirmed that the purified striped bass PRL was also bioactive.
通过二硫键还原和交联研究表征条纹鲈鱼生长激素受体。
DOI：
10.1002/jez.1402680603
发表时间：
1994
期刊：
The Journal of experimental zoology
影响因子：
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作者：
Gray,ES;Tsai,RW
通讯作者：
Tsai,RW