当前位置：
X-MOL 学术
›
Biochemistry
›
论文详情
Our official English website, www.x-mol.net, welcomes your
feedback! (Note: you will need to create a separate account there.)
Characterization of Hsp90 Co-Chaperone p23 Cleavage by Caspase-7 Uncovers a Peptidase–Substrate Interaction Involving Intrinsically Disordered Regions
Biochemistry ( IF 2.9 ) Pub Date : 2017-09-18 00:00:00 , DOI: 10.1021/acs.biochem.7b00298 Cyrielle Martini 1 , Mikaël Bédard 1 , Pierre Lavigne 1 , Jean-Bernard Denault 1
Biochemistry ( IF 2.9 ) Pub Date : 2017-09-18 00:00:00 , DOI: 10.1021/acs.biochem.7b00298 Cyrielle Martini 1 , Mikaël Bédard 1 , Pierre Lavigne 1 , Jean-Bernard Denault 1
Affiliation
|
Caspases are cysteinyl peptidases involved in inflammation and apoptosis during which hundreds of proteins are cleaved by executioner caspase-3 and -7. Despite the fact that caspase-3 has a higher catalytic activity, caspase-7 is more proficient at cleaving poly(ADP ribose) polymerase 1 (PARP1) because it uses an exosite within its N-terminal domain (NTD). Here, we demonstrate that molecular determinants also located in the NTD enhance the recognition and proteolysis of the Hsp90 co-chaperone p23. Structure–activity relationship analyses using mutagenesis of the caspase-7 NTD and kinetics show that residues 36–45 of caspase-7, which overlap with residues necessary for efficacious PARP1 cleavage, participate in p23 recognition. We also demonstrate using chimeric and truncated proteins that the caspase-7 NTD binds close to the cleavage site in the C-terminal tail of p23. Moreover, because p23 is cleaved at a site bearing a P4 Pro residue (PEVD142↓G), which is far from the optimal sequence, we tested all residues at that position and found notable differences in the preference of caspase-7 and magnitude of differences between residues compared to the results of studies that have used small peptidic substrate libraries. Finally, bioinformatics shows that the regions we identified in caspase-7 and p23 are intrinsically disordered regions that contain molecular recognition features that permit a transient interaction between these two proteins. In summary, we characterized the binding mode for a caspase that is tailored to the specific recognition and cleavage of a substrate, highlighting the importance of studying the peptidase–substrate pair to understand the modalities of substrate recognition by caspases.
中文翻译：
Caspase-7 Hsp90伴侣蛋白p23裂解的表征揭示了涉及内在无序区域的肽酶-底物相互作用。
胱天蛋白酶是参与炎症和凋亡的半胱氨酸肽酶，在此过程中，数百种蛋白质被execution子手半胱天冬酶-3和-7裂解。尽管caspase-3具有较高的催化活性，但caspase-7却在切割聚ADP核糖聚合酶1（PARP1）方面更为熟练，因为它在其N端结构域（NTD）中使用了外切位点。在这里，我们证明了分子决定簇也位于NTD中，可增强Hsp90伴侣蛋白p23的识别和蛋白水解作用。利用caspase-7 NTD诱变和动力学进行的结构活性关系分析表明，caspase-7残基36–45与有效PARP1裂解所必需的残基重叠，参与p23识别。我们还证明了使用嵌合蛋白和截短蛋白，caspase-7 NTD结合在p23 C末端尾巴的切割位点附近。此外，由于p23在带有P4 Pro残基的位点（PEVD142 ↓G）（距离最佳序列较远），我们测试了该位置的所有残基，与使用小肽底物文库的研究结果相比，发现caspase-7的偏好性和残基之间的差异程度存在显着差异。最后，生物信息学表明，我们在caspase-7和p23中鉴定的区域是内在无序的区域，其包含分子识别功能，允许这两种蛋白质之间的瞬时相互作用。总而言之，我们表征了针对半胱天冬酶的结合模式，该模式适合于底物的特异性识别和裂解，强调了研究肽酶-底物对以了解半胱天冬酶对底物识别方式的重要性。
更新日期：2017-09-18
中文翻译：
Caspase-7 Hsp90伴侣蛋白p23裂解的表征揭示了涉及内在无序区域的肽酶-底物相互作用。
胱天蛋白酶是参与炎症和凋亡的半胱氨酸肽酶，在此过程中，数百种蛋白质被execution子手半胱天冬酶-3和-7裂解。尽管caspase-3具有较高的催化活性，但caspase-7却在切割聚ADP核糖聚合酶1（PARP1）方面更为熟练，因为它在其N端结构域（NTD）中使用了外切位点。在这里，我们证明了分子决定簇也位于NTD中，可增强Hsp90伴侣蛋白p23的识别和蛋白水解作用。利用caspase-7 NTD诱变和动力学进行的结构活性关系分析表明，caspase-7残基36–45与有效PARP1裂解所必需的残基重叠，参与p23识别。我们还证明了使用嵌合蛋白和截短蛋白，caspase-7 NTD结合在p23 C末端尾巴的切割位点附近。此外，由于p23在带有P4 Pro残基的位点（PEVD142 ↓G）（距离最佳序列较远），我们测试了该位置的所有残基，与使用小肽底物文库的研究结果相比，发现caspase-7的偏好性和残基之间的差异程度存在显着差异。最后，生物信息学表明，我们在caspase-7和p23中鉴定的区域是内在无序的区域，其包含分子识别功能，允许这两种蛋白质之间的瞬时相互作用。总而言之，我们表征了针对半胱天冬酶的结合模式，该模式适合于底物的特异性识别和裂解，强调了研究肽酶-底物对以了解半胱天冬酶对底物识别方式的重要性。
京公网安备 11010802027423号