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蛋白质组学分析人类血管生成素相互作用揭示细胞质 PCNA 作为潜在的结合伴侣。

Proteomic Analysis of Human Angiogenin Interactions Reveals Cytoplasmic PCNA as a Putative Binding Partner.

机构信息

Department of Biochemistry and Biotechnology, University of Thessaly , Biopolis, 41500 Larissa, Greece.

Biomedical Sciences Research Center "Alexander Fleming" , Vari 16672, Greece.

出版信息

J Proteome Res. 2017 Oct 6;16(10):3606-3622. doi: 10.1021/acs.jproteome.7b00335. Epub 2017 Sep 8.

Abstract

Human Angiogenin (hAng) is a member of the ribonuclease A superfamily and a potent inducer of neovascularization. Protein interactions of hAng in the nucleus and cytoplasm of the human umbilical vein cell line EA.hy926 have been investigated by mass spectroscopy. Data are available via ProteomeXchange with identifiers PXD006583 and PXD006584. The first gel-free analysis of hAng immunoprecipitates revealed many statistically significant potential hAng-interacting proteins involved in crucial biological pathways. Surprisingly, proliferating cell nuclear antigen (PCNA), was found to be immunoprecipitated with hAng only in the cytoplasm. The hAng-PCNA interaction and colocalization in the specific cellular compartment was validated with immunoprecipitation, immunoblotting, and immunocytochemistry. The results revealed that PCNA is predominantly localized in the cytoplasm, while hAng is distributed both in the nucleus and in the cytoplasm. hAng and PCNA colocalize in the cytoplasm, suggesting that they may interact in this compartment.

摘要

人血管生成素(hAng)是核糖核酸酶 A 超家族的成员,也是一种有效的新血管生成诱导剂。通过质谱法研究了 hAng 在人脐静脉细胞系 EA.hy926 的细胞核和细胞质中的蛋白质相互作用。数据可通过 ProteomeXchange 获取,标识符为 PXD006583 和 PXD006584。hAng 免疫沉淀物的第一次无胶分析揭示了许多参与关键生物学途径的具有统计学意义的潜在 hAng 相互作用蛋白。令人惊讶的是,只有在细胞质中才发现增殖细胞核抗原(PCNA)与 hAng 免疫沉淀。通过免疫沉淀、免疫印迹和免疫细胞化学验证了 hAng-PCNA 相互作用和在特定细胞区室中的共定位。结果表明,PCNA 主要定位于细胞质,而 hAng 分布于细胞核和细胞质。hAng 和 PCNA 在细胞质中共定位,表明它们可能在该区室中相互作用。

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