Prostate specific antigen cleaves parathyroid hormone-related protein in the PTH-like domain: Inactivation of PTHrP-stimulated cAMP accumulation in mouse osteoblasts
Prostate specific antigen cleaves parathyroid hormone-related protein in the PTH-like domain: Inactivation of PTHrP-stimulated cAMP accumulation in mouse osteoblasts
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DOI：
10.1016/s0022-5347(01)65919-6
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发表时间：
1996-08-01
影响因子：
6.6
通讯作者：
Peehl, DM
中科院分区：
文献类型：
--
作者：
Cramer, SD;Chen, ZX;Peehl, DM
目的：研究甲状旁腺激素相关蛋白（PTHrP）是否为前列腺特异性抗原（PSA）的底物，以及PTHrP与PSA切割后的生物活性变化。材料与方法：重组人PTHrP 1-141在体外37℃下进行前列腺特异性抗原裂解，SDS-PAGE分析。对印迹的PSA切割的PTHrP肽片段进行5轮自动氨基末端氨基酸序列分析，以确定PSA的切割位点。小鼠成骨细胞系MC3T3-E1被用来检测PTHrP 1-141的PSA切割是否改变了其刺激cAMP产生的能力。结果：发现前列腺特异性抗原对PTHrP 1-141具有一定的时间依赖性和剂量依赖性。PSA对PTHrP 1-141的切割在考马斯氏染色的丙烯酰胺凝胶上产生片段，其迁移率分别为19.5、17、15和
Purpose: To determine whether parathyroid hormone-related protein (PTHrP) is a substrate of prostate-specific antigen (PSA) and how the biological activity of PTHrP may be altered by cleavage with PSA.Materials and Methods: Prostate-specific antigen cleavage of recombinant human PTHrP 1-141 was conducted in vitro at 37C and analyzed by SDS-PAGE. Five rounds of automated amino-terminal amino acid sequence analysis were performed on blotted PSA-cleaved PTHrP peptide fragments to determine the PSA cleavage sites. The mouse osteoblast cell line MC3T3-E1 was used to test whether PSA cleavage of PTHrP 1-141 altered its ability to stimulate cAMP production.Results: Prostate-specific antigen was found to specifically cleave PTHrP 1-141 in a time- and dose-dependent manner. Cleavage of PTHrP 1-141 by PSA generated fragments on Coomassie-stained acrylamide gels that migrated with mobilities that corresponded to 19.5, 17, 15 and