Differential Inhibition of Signal Peptide Peptidase Family Members by Established γ-Secretase Inhibitors.
Differential Inhibition of Signal Peptide Peptidase Family Members by Established γ-Secretase Inhibitors.
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DOI：
10.1371/journal.pone.0128619
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Golde TE
中科院分区：
文献类型：
--
作者：
Ran Y;Ladd GZ;Ceballos-Diaz C;Jung JI;Greenbaum D;Felsenstein KM;Golde TE
信号肽肽酶（SPPs）是生物医学上重要的蛋白酶，是丙型肝炎（人SPP， （hSPP））、疟原虫（疟原虫SPP (pSPP)）和b细胞免疫调节和肿瘤（信号肽肽酶如2a， （SPPL2a））的治疗靶点。到目前为止，还没有类似药物的选择性抑制剂的报道。我们使用基于BRI2的氨基端融合到淀粉样蛋白β1-25 (a - β1-25) （FBA）的重组底物来开发简单，成本效益高的SPP/SPPL蛋白酶分析。将表达FBA底物的表达质粒与SPP/SPPLs共转染，通过ELISA、Western Blot和免疫沉淀/MALDI-TOF质谱（IP/MS）监测裂解情况。在没有SPP/SPPL过表达的情况下，未检测到卵裂。多种γ-分泌酶抑制剂（GSIs）和（Z-LL）2酮对SPP/SPPL活性的抑制存在差异；例如，LY-411,575的IC50从51±79 nM （SPPL2a）到5499±122 nM （SPPL2b）不等，而化合物E仅对hSPP有抑制作用，IC50为1465±93 nM。生成的数据预测了小鼠b细胞系中内源性SPPL2a切割CD74的影响。因此，有可能对SPP家族成员进行差异抑制。这些SPP/SPPL裂解试验将加速寻找选择性抑制剂。这些数据也加强了SPP家族成员裂解与γ-分泌酶催化的裂解之间的相似性。
The signal peptide peptidases (SPPs) are biomedically important proteases implicated as therapeutic targets for hepatitis C (human SPP, (hSPP)), plasmodium (Plasmodium SPP (pSPP)), and B-cell immunomodulation and neoplasia (signal peptide peptidase like 2a, (SPPL2a)). To date, no drug-like, selective inhibitors have been reported. We use a recombinant substrate based on the amino-terminus of BRI2 fused to amyloid β 1-25 (Aβ1-25) (FBA) to develop facile, cost-effective SPP/SPPL protease assays. Co-transfection of expression plasmids expressing the FBA substrate with SPP/SPPLs were conducted to evaluate cleavage, which was monitored by ELISA, Western Blot and immunoprecipitation/MALDI-TOF Mass spectrometry (IP/MS). No cleavage is detected in the absence of SPP/SPPL overexpression. Multiple γ-secretase inhibitors (GSIs) and (Z-LL)2 ketone differentially inhibited SPP/SPPL activity; for example, IC50 of LY-411,575 varied from 51±79 nM (on SPPL2a) to 5499±122 nM (on SPPL2b), while Compound E showed inhibition only on hSPP with IC50 of 1465±93 nM. Data generated were predictive of effects observed for endogenous SPPL2a cleavage of CD74 in a murine B-Cell line. Thus, it is possible to differentially inhibit SPP family members. These SPP/SPPL cleavage assays will expedite the search for selective inhibitors. The data also reinforce similarities between SPP family member cleavage and cleavage catalyzed by γ-secretase.
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影响因子：
56.9
作者：
Choudhary, Chunaram;Kumar, Chanchal;Mann, Matthias
通讯作者：
Mann, Matthias
影响因子：
4
作者：
Jonas, Mary Cabell;Pehar, Mariana;Puglielli, Luigi
通讯作者：
Puglielli, Luigi
影响因子：
21.3
作者：
Fluhrer, Regina;Grammer, Gudula;Haass, Christian
通讯作者：
Haass, Christian
影响因子：
3.7
作者：
Hussy, P;Langen, H;Jacobsen, H
通讯作者：
Jacobsen, H
影响因子：
5.4
作者：
Heimann, M;Roman-Sosa, G;Rümenapf, T
通讯作者：
Rümenapf, T