从牛海绵体组织分离纯化磷酸二酯酶5

Identification and Characterization of PDE5 form Bovine Corpus Cavernosum

目的 :纯化磷酸二酯酶 5 (PDE5 )用以研究其抑制剂及临床意义。方法 :利用快速蛋白液相层析系统 ,NaCl线性梯度分离PDE5。结果 :从牛海绵体组织与牛海绵体血管均得到 3个蛋白峰 ,Western印迹实验证实峰 1为PDE5 ,其余二峰可能为PDE2和PDE3。结论 :哺乳类海绵体组织有cGMP特异性PDE5存在 ,在海绵体血管尤为丰富。采用快速蛋白液相系统成功分离PDE5 ,方法简便可靠。

Objective: To isolate PDE5 from bovine related tissues for investigation of pharmacology. Methods: Different PDEs were isolated by FPLC(fast protein liquid chromatography,FPLC) system with a linear NaCl gradient (0～800 mmolL -1 ,60 ml) at 280nm. And PDE5 was identified by western blot analysis. Results: There were 3 peaks both in bovine penis corpus cavernosum tissue and arteries. That peak 1 was identified as PDE5 by western blot assay. The other two may be PDE2 and PDE3. Conclusion: The cGMP-specific cyclic nucleotide phosphodiesterase 5(PDE5) was mainly found in corpus cavernosum tissue of mammalia,especially rich in corpus cavernosum arteries. The hydrolic activity of cGMP in penis was mainly due to PDE5. Using FPLC system to separate PDE5 was convenient and reliable.