Markosian K A, Paĭtian N A, Abramian K S, Nalbandian R M
Biokhimiia. 1989 Dec;54(12):2046-53.
About 40-60% of the peptidylglycine alpha-amidating amonooxygenase activity in the lysates of secretory granules from bovine atria and adrenal medulla isolated and lyzed in the presence of pepstatin, phenylmethylsulfonyl gluoride, N-ethylmaleimide and catalase, was found to be in the soluble form. The remaining part bound to the membrane fraction was extracted with Triton X-100. The procedure of purification of the soluble form of peptidylglycine alpha-amidating monooxygenase from both atrial and chromaffin granules in electrophoretically homogeneous enzyme preparations was developed. The enzyme is made up of a single subunit with a molecular mass of 68 kDa and contains one copper atom per molecule. The EPR spectra of peptidylglycine alpha-amidating amonooxygenase and dopamine beta-monooxygenase were found to be practically identical, thus indicating that the copper environment in the both enzymes is the same. Both peptidylglycine alpha-amidating monooxygenase and dopamine beta-monooxygenase are inhibited by the neurocuprein apoform, an extremely acidic protein isolated from brain and secretory granules of different endocrine tissues.
在胃蛋白酶抑制剂、苯甲基磺酰氟、N-乙基马来酰亚胺和过氧化氢酶存在的情况下，对分离并裂解的牛心房和肾上腺髓质分泌颗粒裂解物中的肽基甘氨酸α-酰胺化单加氧酶活性进行检测，发现约40%-60%的活性呈可溶形式。与膜部分结合的其余部分则用 Triton X-100进行提取。我们开发了从心房颗粒和嗜铬颗粒中纯化肽基甘氨酸α-酰胺化单加氧酶可溶形式的方法，以制备电泳纯的酶制剂。该酶由一个分子量为68 kDa的亚基组成，每个分子含有一个铜原子。发现肽基甘氨酸α-酰胺化单加氧酶和多巴胺β-单加氧酶的电子顺磁共振光谱几乎相同，这表明两种酶中的铜环境相同。肽基甘氨酸α-酰胺化单加氧酶和多巴胺β-单加氧酶都受到神经铜蛋白脱辅基形式的抑制，神经铜蛋白脱辅基形式是一种从不同内分泌组织的脑和分泌颗粒中分离出的极端酸性蛋白质。
