Gibson T R, Zyskind A D, Glembotski C C
Molecular Biology Institute, San Diego State University, California 92182.
J Neurosci. 1988 Aug;8(8):3067-73. doi: 10.1523/JNEUROSCI.08-08-03067.1988.
Atrial natriuretic peptide (ANP) receptors from A10 cultured vascular smooth muscle cells (VSMC) and rat olfactory bulbs have been solubilized and then pharmacologically and biochemically compared. The dissociation constant for 125I-ANP(99-126) was 12.7 pM for the VSMC-derived receptor and 164 pM for the olfactory receptor. Competition binding between 125I-ANP(99-126) and several unlabeled ANP analogs with the soluble olfactory receptor, demonstrated a rank order potency of ANP(99-126) = ANP(103-126) much greater than ANP(103-123). However, the rank order potency of the soluble VSMC ANP receptor was ANP(99-126) = ANP(103-126) = ANP(103-123). Therefore, the olfactory ANP receptor appears to require the complete COOH-terminal sequence of ANP as compared with the VSMC ANP receptor. When the 2 soluble receptor preparations were applied to a GTP-agarose column, a portion of the olfactory ANP receptor was retained on the column and could be eluted with 5 mM GTP, while the VSMC ANP receptor did not adsorb to the column. Since the olfactory bulb ANP receptor has been shown to contain a binding component of 116 kDa, while the VSMC ANP receptor binding component is 66 kDa, these receptors appear to be similar to the 2 receptor classes described recently in which the 120 kDa receptor that binds GTP is postulated to be coupled to guanylate cyclase, while the 60 kDa receptor does not bind GTP, is not coupled to guanylate cyclase, and may possess a hormone clearance function. Taken together, these data indicate that cyclic GMP appears to be a second messenger for ANP in the brain.
已将来自A10培养的血管平滑肌细胞（VSMC）和大鼠嗅球的心房利钠肽（ANP）受体溶解，然后进行药理学和生物化学比较。VSMC衍生受体对125I-ANP(99-126)的解离常数为12.7 pM，嗅球受体的解离常数为164 pM。125I-ANP(99-126)与几种未标记的ANP类似物与可溶性嗅球受体之间的竞争结合表明，ANP(99-126) = ANP(103-126)的效价顺序远大于ANP(103-123)。然而，可溶性VSMC ANP受体的效价顺序为ANP(99-126) = ANP(103-126) = ANP(103-123)。因此，与VSMC ANP受体相比，嗅球ANP受体似乎需要ANP完整的COOH末端序列。当将这两种可溶性受体制剂应用于GTP-琼脂糖柱时，一部分嗅球ANP受体保留在柱上，可用5 mM GTP洗脱，而VSMC ANP受体不吸附到柱上。由于已证明嗅球ANP受体含有116 kDa的结合成分，而VSMC ANP受体结合成分是66 kDa，这些受体似乎类似于最近描述的两类受体，其中与GTP结合的120 kDa受体被认为与鸟苷酸环化酶偶联，而60 kDa受体不与GTP结合，不与鸟苷酸环化酶偶联，可能具有激素清除功能。综上所述，这些数据表明环磷酸鸟苷似乎是大脑中ANP的第二信使。
