Cathepsin L plays a major role in cholecystokinin production in mouse brain cortex and in pituitary AtT-20 cells: protease gene knockout and inhibitor studies.
Cathepsin L plays a major role in cholecystokinin production in mouse brain cortex and in pituitary AtT-20 cells: protease gene knockout and inhibitor studies.
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DOI：
10.1016/j.peptides.2009.06.030
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发表时间：
2009-10
期刊：
影响因子：
3
通讯作者：
Hook V
中科院分区：
文献类型：
--
作者：
Beinfeld MC;Funkelstein L;Foulon T;Cadel S;Kitagawa K;Toneff T;Reinheckel T;Peters C;Hook V
胆囊收缩素 (CCK) 是一种肽类神经递质，其产生需要对 proCCK 前体进行蛋白水解加工，以在大脑中产生活性 CCK8 神经肽。这项研究证明了半胱氨酸蛋白酶组织蛋白酶 L 对 CCK8 生产的重要作用。在组织蛋白酶 L 敲除 (KO) 小鼠中，大脑皮层中的 CCK8 水平显着降低，平均降低了 75%。为了评估组织蛋白酶 L 在神经内分泌细胞调节分泌途径中产生 CCK 的作用，用特异性组织蛋白酶 L 抑制剂 CLIK-148 处理稳定产生 CCK 的垂体 AtT-20 细胞。 CLIK-148 抑制剂治疗导致 AtT-20 细胞受调节的分泌途径分泌的 CCK 量减少。 CLIK-148 还降低了 CCK9 (Arg-CCK8) 的细胞水平，这与 CCK9 作为组织蛋白酶 L 的中间产物一致，如 CCK9/CCK8 比率降低所示。 CCK0/CCK8 比率降低还表明在组织蛋白酶 L 抑制过程中，CCK8 的产生转变为 CCK9。在 siRNA 减少 PC1/3 加工酶的过程中，CCK9/CCK8 的比率增加，表明 CCK9 的产生转变为组织蛋白酶 L 途径。 CCK9 与 CCK8 比率的变化与组织蛋白酶 L 蛋白酶途径（包括去除 NH2 末端 Arg 或 Lys 的氨肽酶 B）和 PC1/3 蛋白酶途径的双重作用一致。这些结果表明，组织蛋白酶 L 作为负责小鼠大脑皮层中 CCK8 产生的主要蛋白酶，并与 PC1/3 一起参与垂体细胞中 CCK8 的产生。
Cholecystokinin (CCK) is a peptide neurotransmitter whose production requires proteolytic processing of the proCCK precursor to generate active CCK8 neuropeptide in brain. This study demonstrates the significant role of the cysteine protease cathepsin L for CCK8 production. In cathepsin L knockout (KO) mice, CCK8 levels were substantially reduced in brain cortex by an average of 75%. To evaluate the role of cathepsin L in producing CCK in the regulated secretory pathway of neuroendocrine cells, pituitary AtT-20 cells that stably produce CCK were treated with the specific cathepsin L inhibitor, CLIK-148. CLIK-148 inhibitor treatment resulted in decreased amounts of CCK secreted from the regulated secretory pathway of AtT-20 cells. CLIK-148 also reduced cellular levels of CCK9 (Arg-CCK8), consistent with CCK9 as an intermediate product of cathepsin L, shown by the decreased ratio of CCK9/CCK8. The decreased CCK0/CCK8 ratio also suggests a shift in the production to CCK8 over CCK9 during inhibition of cathepsin L. During reduction of the PC1/3 processing enzyme by siRNA, the ratio of CCK9/CCK8 was increased, suggesting a shift to the cathepsin L pathway for production of CCK9. The changes in ratios of CCK9 compared to CCK8 are consistent with dual roles of the cathepsin L protease pathway that includes aminopeptidase B to remove NH2-terminal Arg or Lys, and the PC1/3 protease pathway. These results suggest that cathepsin L functions as a major protease responsible for CCK8 production in mouse brain cortex, and participates with PC1/3 for CCK8 production in pituitary cells.
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影响因子：
3
作者：
Beinfeld, MC;Vishnuvardhan, D;Marchand, JE
通讯作者：
Marchand, JE
影响因子：
3.9
作者：
Katunuma, N;Tsuge, H;Fukushima, M
通讯作者：
Fukushima, M
DOI：
10.1073/pnas.90.14.6691
发表时间：
1993-07-15
影响因子：
11.1
作者：
LUSSON, J;VIEAU, D;SEIDAH, NG
通讯作者：
SEIDAH, NG
影响因子：
4.1
作者：
CADEL, S;PIEROTTI, AR;COHEN, P
通讯作者：
COHEN, P
影响因子：
3.6
作者：
Kitagawa, K;Aida, C;Inoue, K
通讯作者：
Inoue, K