Lys40影响血管紧张素转化α-酶的选择性,而Arg143不影响

Diego J. Muilenburg, Wilfred W. Raymond, Paul J. Wolters, George H. Caughey
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引用次数: 26

摘要

人α-切酶是一种高效的血管紧张素(AT)转化酶,在Phe8下选择性水解AT I,生成具有生物活性的AT II,可促进心脏肥厚、血管狭窄和高血压。一些相关的酶,如大鼠β-切酶1,选择性低得多,通过在Tyr4上切割来破坏AT。通过对酶结构和活性的比较,我们推测AT与Lys40或Arg143侧链之间的相互作用是人类酶对Phe8的明显偏好超过Tyr4的原因。为了验证这些假设,我们比较了野生型切酶和将Lys40或Arg143转变为中性残基的突变体对AT的水解。Lys40被犬α-和大鼠β-切酶1中发现的残基丙氨酸所交换,后者在Phe8时的水解选择性明显降低。用Arg143与大鼠β-切酶1中的谷氨酰胺交换。Lys40Ala突变体是一种类似狗的酶,保留了对Phe8的强烈偏好,但与野生型人酶相比,Tyr4水解率提高了16倍。因此,在狗和人酶之间的40个不匹配的残基中,单个残基占了它们之间特异性差异的大部分。与预测相反,Arg143Gln突变体仍然具有高度的phe8选择性。因此,Lys40,而不是Arg143,促成了人类乳糜酶对AT II生成而不是破坏的显著偏好。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Lys40 but not Arg143 influences selectivity of angiotensin conversion by human α-chymase

Human α-chymase is an efficient angiotensin (AT) converting enzyme, selectively hydrolyzing AT I at Phe8 to generate bioactive AT II, which can promote cardiac hypertrophy, vascular stenosis, and hypertension. Some related enzymes, such as rat β-chymase 1, are much less selective, destroying AT by cleaving at Tyr4. Comparisons of chymase structure and activity led to speculation that interaction between AT and the side chain of Lys40 or Arg143 accounts for the human enzyme’s marked preference for Phe8 over Tyr4. To test these hypotheses, we compared AT hydrolysis by wild-type chymase with that by mutants changing Lys40 or Arg143 to neutral residues. Lys40 was exchanged for alanine, the residue found in canine α- and rat β-chymase 1, the latter being dramatically less selective for hydrolysis at Phe8. Arg143 was exchanged for glutamine found in rat β-chymase 1. The Lys40Ala mutant is a dog-like enzyme retaining strong preference for Phe8 but with Tyr4 hydrolytic rates enhanced 16-fold compared to wild-type human enzyme. Thus, of 40 residues mismatched between dog and human enzymes, a single residue accounts for most of the difference in specificity between them. The Arg143Gln mutant, contrary to prediction, remains highly Phe8-selective. Therefore, Lys40, but not Arg143, contributes to human chymase’s remarkable preference for AT II generation over destruction.

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