MOLECULAR AND BIOCHEMICAL-CHARACTERIZATION OF THE MOUSE-BRAIN CORTICOTROPIN-RELEASING HORMONE-BINDING PROTEIN
MOLECULAR AND BIOCHEMICAL-CHARACTERIZATION OF THE MOUSE-BRAIN CORTICOTROPIN-RELEASING HORMONE-BINDING PROTEIN
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DOI：
10.1016/0303-7207(95)03558-o
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发表时间：
1995-06-01
影响因子：
4.1
通讯作者：
SEASHOLTZ, AF
中科院分区：
文献类型：
--
作者：
CORTRIGHT, DN;NICOLETTI, A;SEASHOLTZ, AF
与 CRH 受体不同的 37 kDA 促肾上腺皮质激素释放激素结合蛋白 (CRH-BP) 在大鼠垂体前叶促肾上腺皮质激素和大脑的许多区域中表达，表明 CRH-BP 可能调节 CRH 的生物活性。在这些研究中，小鼠大脑 CRH-BP (mCRH-BP) cDNA 已被分离和表征。 1666 个核苷酸的 mCRH-BP cDNA 在大脑和垂体中表达，编码与人和大鼠 CRH-BP 高度同源的 322 个氨基酸的蛋白质。在培养的哺乳动物细胞中表达的重组 mCRH-BP 以高亲和力结合人 CRH（K-d(app)= 0.56 nM 和 K-i(app)= 0.37 nM）和 α-螺旋 (9-41) CRH 拮抗剂（Ki(app)= 0.28 nM），但对绵羊 CRH 的亲和力要弱得多（K-i(app)= 206 nM）。重组 mCRH-BP 还可阻断 CRH 诱导的 AYT-20 细胞释放促肾上腺皮质激素。 mCRH-BP 结合活性的其他生化特征表明 hRH-BP 和 CRH 受体利用不同的分子相互作用来结合 CRH。
A 37 kDA corticotropin-releasing hormone-binding protein (CRH-BP), distinct from the CRH receptor, is expressed in rat anterior pituitary corticotrophs and many regions of the brain, suggesting that CRH-BP may modulate the biological activity of CRH. In these studies a mouse brain CRH-BP (mCRH-BP) cDNA has been isolated and characterized. The 1666 nucleotide mCRH-BP cDNA is expressed in brain and pituitary and encodes a 322 amino acid protein that is highly homologous to human and rat CRH-BPs. Recombinant mCRH-BP, expressed in cultured mammalian cells, binds human CRH (K-d(app)= 0.56 nM and K-i(app)= 0.37 nM) and the alpha-helical (9-41) CRH antagonist (Ki(app)= 0.28 nM) with high affinity, but exhibits much weaker affinity for ovine CRH (K-i(app)= 206 nM). Recombinant mCRH-BP also blocks CRH-induced adrenocorticotropin release from AYT-20 cells. Additional biochemical characterization of the binding activity of mCRH-BP indicates that hRH-BP and CRH receptor utilize different molecular interactions to bind CRH.