Beaumont A, Roques B P
Biochem Biophys Res Commun. 1986 Sep 14;139(2):733-9. doi: 10.1016/s0006-291x(86)80052-3.
Diethylpyrocarbonate treatment of the neutral endopeptidase (EC 3.4.24.11) inhibits both catalytic activity and binding of the inhibitor [3H]-N(R,S)-3-hydroxyaminocarbonyl-2-benzyl-1-oxopropyl]-glycine. The loss of activity can be reversed by hydroxylamine and almost completely prevented by the competitive inhibitor phenylalanyl-leucine suggesting the presence, as in thermolysin, of a histidine residue at the active site. Butanedione treatment also reduces both catalytic activity and [3H] inhibitor binding. Phenylalanyl-leucine completely protects from the butanedione induced loss of activity, providing further evidence for an essential arginine at the active site. In contrast, the tyrosine modifying agent N-acetylimidazole has no apparent effect on enzyme activity.
焦碳酸二乙酯处理中性内肽酶（EC 3.4.24.11）会抑制其催化活性以及抑制剂[3H]-N（R，S）-3-羟基氨基羰基-2-苄基-1-氧代丙基]-甘氨酸的结合。活性的丧失可通过羟胺逆转，并且几乎完全被竞争性抑制剂苯丙氨酰-亮氨酸阻止，这表明如同嗜热菌蛋白酶一样，活性位点存在组氨酸残基。丁二酮处理也会降低催化活性和[3H]抑制剂结合。苯丙氨酰-亮氨酸完全保护酶免受丁二酮诱导的活性丧失，为活性位点存在必需的精氨酸提供了进一步证据。相比之下，酪氨酸修饰剂N-乙酰咪唑对酶活性没有明显影响。
