Lipid environment modulates processivity and kinetics of a presenilin homolog acting on multiple substrates in vitro.
Lipid environment modulates processivity and kinetics of a presenilin homolog acting on multiple substrates in vitro.
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DOI：
10.1016/j.jbc.2023.105401
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发表时间：
2023-12
影响因子：
4.8
通讯作者：
Lieberman, Raquel L.
中科院分区：
文献类型：
--
作者：
Wu, Yuqi;Thomas, Gwendell M.;Thomsen, Max;Bahri, Sara;Lieberman, Raquel L.
膜内蛋白酶（IPs）水解脂质膜中的肽。IPs参与许多细胞途径，包括免疫反应和监视，胆固醇生物合成，它们被病毒利用进行复制。尽管它们在生物学中具有广泛的重要性，但细胞中如何调节活性以控制蛋白质成熟和在正确的时间和地点释放特定生物活性肽仍然没有很大的答案，特别是对于膜内天冬氨酸蛋白酶（IAP）亚型。在分子生化水平上，不同的IAP同源物可以切割非生物底物，并且在近150个底物中没有序列识别基序，只有一个IAP，即早老素-1，γ分泌酶的催化成分，以其参与与阿尔茨海默病相关的淀粉样蛋白-β斑块的产生而知名。在这里，我们使用凝胶为基础的分析结合定量质谱和基于fret的动力学分析来探测甲烷菌Methanoculleus marisnigri JR1的早老素同源物的裂解谱，作为周围脂质模拟环境（洗涤剂胶束或单束）的函数。我们选择了四种生物IAP底物，即丙型肝炎病毒的病毒核心蛋白、经典猪瘟病毒的病毒核心蛋白、Notch-1的跨膜片段和酪氨酸受体激酶ErbB4。我们的研究证明了底物在低平均疏水性位置的裂解倾向，以及脂质环境在调节动力学性质中的一致作用。
Intramembrane proteases (IPs) hydrolyze peptides in the lipid membrane. IPs participate in a number of cellular pathways including immune response and surveillance, and cholesterol biosynthesis, and they are exploited by viruses for replication. Despite their broad importance across biology, how activity is regulated in the cell to control protein maturation and release of specific bioactive peptides at the right place and right time remains largely unanswered, particularly for the intramembrane aspartyl protease (IAP) subtype. At a molecular biochemical level, different IAP homologs can cleave non-biological substrates, and there is no sequence recognition motif among the nearly 150 substrates identified for just one IAP, presenilin-1, the catalytic component of γ-secretase known for its involvement in the production of amyloid-β plaques associated with Alzheimer disease. Here we used gel-based assays combined with quantitative mass spectrometry and FRET-based kinetics assays to probe the cleavage profile of the presenilin homolog from the methanogen Methanoculleus marisnigri JR1 as a function of the surrounding lipid-mimicking environment, either detergent micelles or bicelles. We selected four biological IAP substrates that have not undergone extensive cleavage profiling previously, namely, the viral core protein of Hepatitis C virus, the viral core protein of Classical Swine Fever virus, the transmembrane segment of Notch-1, and the tyrosine receptor kinase ErbB4. Our study demonstrates a proclivity toward cleavage of substrates at positions of low average hydrophobicity and a consistent role for the lipid environment in modulating kinetic properties.
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影响因子：
7.7
作者：
Feilen, Lukas P.;Chen, Shu-Yu;Fukumori, Akio;Feederle, Regina;Zacharias, Martin;Steiner, Harald
通讯作者：
Steiner, Harald
影响因子：
11.4
作者：
Arutyunova, Elena;Panwar, Pankaj;Lemieux, M. Joanne
通讯作者：
Lemieux, M. Joanne
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