通过叠氮化物介导的交联作用靶向 parvulin 相互作用蛋白，揭示了人类 Par14/17 在肌动蛋白聚合中的细胞作用。

Targeting of parvulin interactors by diazirine mediated cross-linking discloses a cellular role of human Par14/17 in actin polymerization.

机构信息

Structural and Medicinal Biochemistry, Center for Medical Biotechnology (ZMB), University of Duisburg-Essen, Universitätsstrasse 2-5, D-45117 Essen, Germany.

Imaging Centre Campus Essen (ICCE), Center for Medical Biotechnology (ZMB), University of Duisburg-Essen, Universitätsstrasse 2-5, D-45117 Essen, Germany.

出版信息

Biol Chem. 2020 Jul 28;401(8):955-968. doi: 10.1515/hsz-2019-0423.

DOI:10.1515/hsz-2019-0423

PMID:32142471

Abstract

The peptidyl-prolyl cis/trans isomerases (PPIases) Parvulin 14 (Par14) and Parvulin 17 (Par17) result from alternative transcription initiation of the PIN4 gene. Whereas Par14 is present in all metazoan, Par17 is only expressed in Hominidae. Par14 resides mainly within the cellular nucleus, while Par17 is translocated into mitochondria. Using photo-affinity labeling, cross-linking and mass spectrometry (MS) we identified binding partners for both enzymes from HeLa lysates and disentangled their cellular roles. Par14 is involved in biogenesis of ribonucleoprotein (RNP)-complexes, RNA processing and DNA repair. Its elongated isoform Par17 participates in protein transport/translocation and in cytoskeleton organization. Nuclear magnetic resonance (NMR) spectroscopy reveals that Par17 binds to β-actin with its N-terminal region, while both parvulins initiate actin polymerization depending on their PPIase activity as monitored by fluorescence spectroscopy. The knockdown (KD) of Par17 in HCT116 cells results in a defect in cell motility and migration.

摘要

肽基脯氨酰顺/反式异构酶（PPIases）Parvulin 14（Par14）和 Parvulin 17（Par17）是由 PIN4 基因的转录起始的替代产生的。Par14 存在于所有后生动物中，而 Par17 仅在人科中表达。Par14 主要位于细胞核内，而 Par17 则被转运到线粒体中。通过光亲和标记、交联和质谱（MS）分析，我们从 HeLa 裂解物中鉴定出两种酶的结合伴侣，并阐明了它们的细胞功能。Par14 参与核糖核蛋白（RNP）复合物的生物发生、RNA 加工和 DNA 修复。其延伸异构体 Par17 参与蛋白质运输/易位和细胞骨架组织。核磁共振（NMR）光谱表明 Par17 与 β-肌动蛋白结合在其 N 端区域，而这两种 parvulin 都依赖于其 PPIase 活性启动肌动蛋白聚合，这可以通过荧光光谱监测到。在 HCT116 细胞中敲低 Par17 会导致细胞迁移和运动缺陷。