Detection of prolactin based on the receptor-binding activity and its application

基于受体结合活性的催乳素检测及其应用

• 批准号: 02556037
• 负责人: SAKAI Senkiti
• 金额: $ 3.01万
• 依托单位: The university of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research (B)
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1991
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02556037/
• 关键词: prolactin; radioreceptorassay; radioimmunoasiay; receptor; immobilized-receptor; ラジオレセプタ-アッセ-; プロラクチンレセプタ-; 標識プロラクチン

Prolactin sends its hormonal message to target organs through binding with its receptor located on the plasma membrane. Shill et al. reported that the receptor-binding activity of prolactin correlates well with its biological activity. -However, various molecular forms of prolactin have been identified. Among them, for example glycosylatede prolactin is less active than nonglycosylated prolactin. The microsome fraction has been used as a source of the receptor. In the present experiments, we solubilized these membrane proteins with Triton X-100, purified partially using Con-A affinity chromatography, and then immobilized them on a nitrocellulose membrane filter. Using membrane-bound receptor and bovine serum, the serum level of prolactin was determined by radioreceptor assay. The dosedisplacement curve was obtained in the range of 0.4 and 20 ng/ml. Serum could be included in concentrations up to 16% of the assay buffer and, thus the detection limit was about 2.5 ng/ml. Prolactin in the serum was determined by mdioimmunoassay. The ratio of the two assays was nearly 0.99 (r>+O. 99, n=55). The gel filtration profile was identical between the two assays. It was concluded that membranebound receptor can be used for the determination of serum prolactin with satisfactory.

催乳素通过与位于质膜上的受体结合将其激素信息发送到靶器官。Shill等人报道，催乳素的受体结合活性与其生物活性密切相关。然而，催乳素的各种分子形式已被鉴定。其中，例如糖基化催乳素的活性低于非糖基化催乳素。微粒体部分已被用作受体的来源。在本实验中，我们用Triton X-100溶解这些膜蛋白，用Con-A亲和层析部分纯化，然后将它们固定在硝酸纤维素膜过滤器上。用放射受体法测定血清催乳素水平。在0.4 ~ 20 ng/ml范围内获得了剂量位移曲线。血清可以以高达测定缓冲液的16%的浓度包含，因此检测限为约2.5 ng/ml。血清催乳素测定采用间接免疫法。两种方法的比值接近0.99（r>+0. 99，n=55）。两种试验的凝胶过滤曲线相同。结果表明，膜结合受体可用于血清催乳素的测定。

项目成果

Sakai, S: "Effect of hormone on dissociation form the rabbit mammary gland prolactin receptor." Biochemical Journal. 279. 461-465 (1991)

Sakai，S：“激素对兔乳腺催乳素受体解离的影响。”

SUZUKI,M.,KOHMOTO,K.,SAKAI,S: "Radio receptor assay of serum prolactin using nitrocellulose memlerne-immobilized mammary prolactin receptor" Analytical Biochemistry. 200. 42-46 (1992)

SUZUKI,M.,KOHMOTO,K.,SAKAI,S：“使用硝化纤维素膜固定的乳腺催乳素受体对血清催乳素进行放射受体测定”分析生物化学。

SUZUKI,M.,KOHMOTO.K.,SAKAI,S: "Radioreceptor assay of serum prolactin using nitrocellulose memhrne-immobilized mammary prolactin receptor" Analytical Biochemistry. 200. 42-46 (1992)

SUZUKI,M.,KOHMOTO.K.,SAKAI,S：“使用硝化纤维素膜固定的乳腺催乳素受体对血清催乳素进行放射受体测定”分析生物化学。

Sakai, S., Yamamoto, K. and Nagasawa, H.: "Prolacting and progesterone receptors in pregnancy-dependent mammary tumors in GR/A mice." Proceeding of the Society for Experimental Biology and Medicine. 195. 375-378 (1990)

Sakai, S.、Yamamoto, K. 和 Nagasawa, H.：“GR/A 小鼠妊娠依赖性乳腺肿瘤中的催乳素和黄体酮受体。”

SAKAI,S.,SUZUKI,M.KOHMOTO,K: "Thermodynemic analysis of the interaction of prolactin with its receptor in the rabbit mammary-gland miassones" Biochemical Journal. 269. 647-650 (1990)

SAKAI,S.,SUZUKI,M.KOHMOTO,K：“催乳素与其受体在兔乳腺 miassones 中相互作用的热力学分析”生物化学杂志。