Peptidic phosphonylating agents as irreversible inhibitors of serine proteases and models of the tetrahedral intermediates.
Peptidic phosphonylating agents as irreversible inhibitors of serine proteases and models of the tetrahedral intermediates.
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DOI：
10.1021/bi00222a031
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发表时间：
1991-02
期刊：
影响因子：
2.9
通讯作者：
N. Sampson;P. Bartlett
中科院分区：
文献类型：
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作者：
N. Sampson;P. Bartlett
已经合成并研究了掺入亲电磷部分（2-6）的肽类似物作为多种丝氨酸蛋白酶的抑制剂。抑制是不可逆的，并且对于α-裂解蛋白酶（ALP），显示是由与活性位点丝氨酸羟基的共价结合引起的[Bone，R.，Sampson，N.美国，巴特利特，宾夕法尼亚州，& Agard，D. A.（1991）生物化学（以下文件在这个问题）]。对于人白细胞弹性蛋白酶（HLE）与噻吩酯6s-V（Boc-AAPV psi [P = O（SPh）O]AA-OMe）、4s-V（BocAAPV psi [P = O（SPh）O]-Me）和3s-V（Boc-V psi [P = O（SPh）O]AA-OMe）的反应，有证据表明在失活过程中抑制剂占据S4-S1亚位点，而不是S1'和S2'位置。在蛋白酶和六肽膦酸酯6 o-V（Boc-AAPV psi [P = O（OPh）O]AA-OMe）和6 o-F（Boc-AAPF psi [P = O（OPh）O]AA-OMe）之间观察到的选择性与这些酶和它们的底物之间的选择性相似：ALP和HLE被含ValP的类似物6 o-V选择性灭活，而枯草杆菌蛋白酶（BHP）显示出对PheP衍生物6 o-F的偏好。酶-抑制剂相互作用的详细动力学分析是复杂的抑制剂酶降解的敏感性。在磷的配置被认为是没有显着的影响，抑制剂与肽酶反应的速率。此外，在六肽6 o-V使ALP失活的情况下，由两种立体异构体形成相同的共价加合物（Bone等人，1991），表明这些非对映异构体中的一种经历取代而保留构型。
Peptide analogues incorporating an electrophilic phosphorus moiety (2-6) have been synthesized and studied as inhibitors of a variety of serine proteases. Inhibition is irreversible and, for alpha-lytic protease (ALP), shown to result from covalent binding to the active site serine hydroxyl [Bone, R., Sampson, N. S., Bartlett, P.A., & Agard, D. A. (1991) Biochemistry (following paper in this issue)]. For reaction of human leukocyte elastase (HLE) with the thiophenyl esters 6s-V (Boc-AAPV psi [P = O(SPh)O]AA-OMe), 4s-V (BocAAPV psi [P = O(SPh)O]-Me), and 3s-V (Boc-V psi [P = O(SPh)O]AA-OMe), evidence is presented to suggest that the S4-S1 subsites, but not the S1' and S2' positions, are occupied by the inhibitors during the inactivation process. The selectivity that is observed between the proteases and the hexapeptide phosphonates 6o-V (Boc-AAPV psi [P = O(OPh)O]AA-OMe) and 6o-F (Boc-AAPF psi [P = O(OPh)O]AA-OMe) parallels that between these enzymes and their substrates: ALP and HLE are selectively inactivated by the ValP-containing analogue 6o-V, while subtilisin (SUB) shows a preference for the PheP derivative 6o-F. A detailed kinetic analysis of the enzyme-inhibitor interactions was complicated by the susceptibility of the inhibitors to enzymatic degradation. The configuration at phosphorus was found not to have a significant influence on the rate at which the inhibitors react with the peptidases. Moreover, in the case of inactivation of ALP by the hexapeptide 6o-V, the same covalent adduct is formed from both stereoisomers (Bone et al., 1991), indicating that one of these diastereomers undergoes substitution with retention of configuration.