Molecular Features of CA-074 pH-Dependent Inhibition of Cathepsin B.
Molecular Features of CA-074 pH-Dependent Inhibition of Cathepsin B.
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DOI：
10.1021/acs.biochem.1c00684
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发表时间：
2022-02-15
期刊：
影响因子：
2.9
通讯作者：
Hook, Vivian
中科院分区：
文献类型：
--
作者：
Yoon, Michael C.;Christy, Mitchell P.;Phan, Von V.;Gerwick, William H.;Hook, Gregory;O'Donoghue, Anthony J.;Hook, Vivian
CA-074是组织蛋白酶B（一种溶酶体半胱氨酸蛋白酶）的选择性抑制剂。CA-074在许多研究中被用来证明这种蛋白酶在细胞和生理功能中的作用。组织蛋白酶B在许多人类疾病机制中涉及其从pH 4.6的酸性溶酶体转移到pH 7.2的细胞位置，包括细胞质和细胞外环境。为了深入了解CA-074在这些不同pH条件下的抑制作用，本研究评估了CA-074在酸性和中性pH条件下对组织蛋白酶B的抑制作用的分子特征、效力和选择性。本研究表明，CA-074在pH为4.6的酸性条件下对组织蛋白酶B的抑制效果最好，效价为nM，比中性pH为7.2时的抑制效果强100倍以上。CA-074的ph依赖性抑制被其c端脯氨酸的甲基化所消除，表明ph依赖性抑制需要游离的c端羧基。在这些酸性和中性的pH条件下，CA-074对组织蛋白酶B保持了对其他半胱氨酸组织蛋白酶的特异性，表现出不可逆的抑制作用，并且通过谱分析质谱法评估了CA-074对组织蛋白酶B肽底物的多种裂解。分子对接表明，CA-074的c端羧酸盐在pH值为4.6时与酶S2'亚位上的His110和His111残基发生pH依赖的离子相互作用，但在pH值为7.2时这些相互作用不同。虽然高水平的CA-074或CA-074Me（通过细胞酯酶转化为CA-074）在生物学研究中被用于在酸性和中性pH值下抑制组织蛋白酶B，但可能会探索调整CA-074或CA-074Me的水平，以不同程度地影响这些不同pH值下的组织蛋白酶B活性。总的来说，本研究的结果证明了CA-074 ph依赖性组织蛋白酶B抑制的分子、动力学和蛋白酶特异性特征。
CA-074 is a selective inhibitor of cathepsin B, a lysosomal cysteine protease. CA-074 has been utilized in numerous studies to demonstrate the role of this protease in cellular and physiological functions. Cathepsin B in numerous human disease mechanisms involves its translocation from acidic lysosomes of pH 4.6 to neutral pH 7.2 of cellular locations including the cytosol and extracellular environment. To gain in-depth knowledge of CA-074 inhibition at these different pH conditions, this study evaluated the molecular features, potency, and selectivity of CA-074 for cathepsin B inhibition at acidic and neutral pH conditions. This study demonstrated that CA-074 is most effective at inhibiting cathepsin B at the acidic pH of 4.6 with nM potency, which was more than 100-fold more potent than its inhibition at the neutral pH of 7.2. The pH-dependent inhibition of CA-074 was abolished by methylation of its C-terminal proline, indicating the requirement for the free C-terminal carboxyl group for pH-dependent inhibition. At these acidic and neutral pH conditions, CA-074 maintained its specificity for cathepsin B over other cysteine cathepsins, displayed irreversible inhibition, and inhibited diverse cleavages of peptide substrates of cathepsin B assessed by profiling mass spectrometry. Molecular docking suggested that pH-dependent ionic interactions of the C-terminal carboxylate of CA-074 occur with His110 and His111 residues in the S2' subsite of the enzyme at pH 4.6, but these interactions differ at pH 7.2. While high levels of CA-074 or CA-074Me (converted by cellular esterases to CA-074) are used in biological studies to inhibit cathepsin B at both acidic and neutral pH locations, it is possible that adjusted levels of CA-074 or CA-074Me may be explored to differentially affect cathepsin B activity at these different pHs. Overall, results of this study demonstrates the molecular, kinetic, and protease specificity features of CA-074 pH-dependent inhibition of cathepsin B.
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影响因子：
4.7
作者：
Jiang M;Meng J;Zeng F;Qing H;Hook G;Hook V;Wu Z;Ni J
通讯作者：
Ni J
组织蛋白酶B促进结直肠肿瘤发生，细胞侵袭和转移。
影响因子：
4.6
作者：
Bian B;Mongrain S;Cagnol S;Langlois MJ;Boulanger J;Bernatchez G;Carrier JC;Boudreau F;Rivard N
通讯作者：
Rivard N
影响因子：
7
作者：
Lapek, John D., Jr.;Jiang, Zhenze;O'Donoghue, Anthony J.
通讯作者：
O'Donoghue, Anthony J.
影响因子：
4.1
作者：
BARRETT, AJ;KEMBHAVI, AA;HANADA, K
通讯作者：
HANADA, K
DOI：
10.1002/prca.201300105
发表时间：
2014-06
期刊：
Proteomics. Clinical applications
影响因子：
--
作者：
Aggarwal N;Sloane BF
通讯作者：
Sloane BF