Characterization of an A-Site Selective Protein Disulfide Isomerase A1 Inhibitor.
Characterization of an A-Site Selective Protein Disulfide Isomerase A1 Inhibitor.
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DOI：
10.1021/acs.biochem.8b00178
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发表时间：
2018-04-03
期刊：
影响因子：
2.9
通讯作者：
Weerapana E
中科院分区：
文献类型：
--
作者：
Cole KS;Grandjean JMD;Chen K;Witt CH;O'Day J;Shoulders MD;Wiseman RL;Weerapana E
蛋白二硫异构酶A1 （PDIA1）是一种内质网（ER）定位的硫醇二硫氧化还原酶，是分泌途径蛋白的重要折叠催化剂。PDIA1包含两个活性位点域（a和a '），每个域都包含一个Cys-Gly-His-Cys （CGHC）活性位点基序。这两个活性位点结构域具有37%的序列同一性，并独立地进行二硫键还原、氧化和异构化。已经报道了许多PDIA1抑制剂，但这些抑制剂对a和a '位点的选择性尚不清楚。在这里，我们发现了一种有效的、选择性的PDIA1抑制剂KSC-34，它对a位点的选择性是a '位点的30倍。KSC-34在体外表现出对PDIA1还原酶活性的时间依赖性抑制，其kinact/KI = 9.66 × 103 M−1s−1，并且对PDIA1具有选择性，而不是PDI家族的其他成员和其他细胞含半胱氨酸蛋白。我们首次提供了a位点选择性PDIA1抑制剂的细胞表征，并证明KSC-34对细胞未折叠蛋白反应（UPR）具有最小的持续影响，这表明a位点抑制不会诱导全局蛋白质折叠相关的内质网应激。KSC-34处理显著减少了不稳定的淀粉样蛋白抗体轻链的分泌，从而最大限度地减少了依赖高PDIA1活性进行适当折叠和分泌的致病性淀粉样蛋白细胞外蛋白。鉴于人们对PDIA1各活性位点对PDIA1病理生理功能的贡献知之甚少，位点选择性抑制剂如KSC-34为描述PDIA1的病理作用和治疗潜力提供了有用的工具。
Protein disulfide isomerase A1 (PDIA1) is an endoplasmic reticulum (ER)-localized thiol-disulfide oxidoreductase that is an important folding catalyst for secretory pathway proteins. PDIA1 contains two active-site domains (a and a′), each containing a Cys-Gly-His-Cys (CGHC) active-site motif. The two active-site domains share 37% sequence identity, and function independently to perform disulfide-bond reduction, oxidation and isomerization. Numerous inhibitors for PDIA1 have been reported, yet the selectivity of these inhibitors toward the a and a′ sites are poorly characterized. Here, we identify a potent and selective PDIA1 inhibitor, KSC-34, with 30-fold selectivity for the a site over the a′ site. KSC-34 displays time-dependent inhibition of PDIA1 reductase activity in vitro with a kinact/KI = 9.66 × 103 M−1s−1, and is selective for PDIA1 over other members of the PDI family, and other cellular cysteine-containing proteins. We provide the first cellular characterization of an a-site selective PDIA1 inhibitor and demonstrate that KSC-34 has minimal sustained effects on the cellular unfolded protein response (UPR), indicating that a-site inhibition does not induce global protein-folding-associated ER stress. KSC-34 treatment significantly decreases secretion of a destabilized, amyloidogenic antibody light chain, thereby minimizing pathogenic amyloidogenic extracellular proteins that rely on high PDIA1 activity for proper folding and secretion. Given the poor understanding of the contribution of each PDIA1 active site to the (patho)physiological functions of PDIA1, site-selective inhibitors like KSC-34 provide useful tools for delineating the pathological role and therapeutic potential of PDIA1.
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影响因子：
16.6
作者：
Eirich, Juergen;Braig, Simone;Vollmar, Angelika M.
通讯作者：
Vollmar, Angelika M.
影响因子：
4.8
作者：
Mayer, M;Kies, U;Buchner, J
通讯作者：
Buchner, J
影响因子：
64.8
作者：
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通讯作者：
Thomson, Elizabeth
DOI：
10.1073/pnas.1500439112
发表时间：
2015-04-28
影响因子：
11.1
作者：
Kaplan, Anna;Gaschler, Michael M.;Stockwell, Brent R.
通讯作者：
Stockwell, Brent R.
影响因子：
11.4
作者：
Klappa, P;Ruddock, LW;Freedman, RB
通讯作者：
Freedman, RB