Separation and identification of hsp27 interacting proteins from human hepatocellular carcinoma cells

SUN Xue-feng; et al

doi:10.13406/j.cnki.cyxb.2010.08.035

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Separation and identification of hsp27 interacting proteins from human hepatocellular carcinoma cells

更新时间：2024-01-17

- Separation and identification of hsp27 interacting proteins from human hepatocellular carcinoma cells
- Journal of Chongqing Medical University Vol. 35, Issue 8, Pages: 1221-1223(2010)
- 作者机构：
  
  重庆医科大学附属第二医院消化内科
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2010.08.035
  CLC： R735.7
- Published：2010，
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[1]孙雪峰,何松.肝癌细胞中热休克蛋白27相互作用蛋白的分离和鉴定[J].重庆医科大学学报,2010,35(08):1221-1223.

SUN Xue-feng, et al. Separation and identification of hsp27 interacting proteins from human hepatocellular carcinoma cells. [J]. Journal of Chongqing Medical University 35(8):1221-1223(2010)
[1]孙雪峰,何松.肝癌细胞中热休克蛋白27相互作用蛋白的分离和鉴定[J].重庆医科大学学报,2010,35(08):1221-1223. DOI： 10.13406/j.cnki.cyxb.2010.08.035.

SUN Xue-feng, et al. Separation and identification of hsp27 interacting proteins from human hepatocellular carcinoma cells. [J]. Journal of Chongqing Medical University 35(8):1221-1223(2010) DOI： 10.13406/j.cnki.cyxb.2010.08.035.

摘要

目的:筛选肝癌中热休克蛋白27（Heat shock protein

HSP27）相互作用蛋白。方法:采用免疫共沉淀技术富集肝癌HepG2.2215细胞的HSP27结合蛋白

直接酶解后进行液相色谱-电喷雾离子阱串联质谱分析（LC-ESI-ITMS/MS）

通过相应的肽序列标签（Peptide sequence tags

PST）在SWISS PROT数据库中搜索蛋白质。并用免疫共沉淀和蛋白质印迹分析技术对所鉴定的HSP27相互作用蛋白之一HSP90进行了验证。结果:通过搜索数据库在肝癌细胞系中首次鉴定了12个HSP27结合蛋白

分别是HSP90、HSP70、特异性下调葡萄糖调节蛋白78（GRP-78）、赖氨酰羟化酶（Lysyl hydroxylase）、细胞角蛋白9（Keratin 9）、细胞角蛋白10（Keratin 10）和细胞角蛋白1（Keratin 1）。所鉴定的7个结合蛋白可以划分为3个作用范畴

分别是:分子伴侣、细胞骨架组织和代谢酶类。结论:HSP27在肝癌组织中参与了相关蛋白的调节

通过与一些多肽或小分子蛋白质的相互作用

而参与调控肿瘤发生发展。

Abstract

Objective:To find out heatshock protein HSP27 interactingproteins in the hepatocellularcarcinoma(HCC).Methods:HSP27 interacting proteins in the human HCC HepG2.2215 cells were seperated and identified.HSP27 binding proteins were recovered by antiHSP27 antibody immunoprecipitation with the total proteins from HCC HepG2.2215 cells.After in-solution digested and analyzed by LC-ESI-ITMS/MS

the recovered protein complexes were identified by peptide sequence tags(PST)and database searching

and were confimed by immunoprecipitation and Western blot analysis.Results:Seven HSP27 binding proteins were firstly identified by database searching

which were HSP90

GRP-78 and HSP70 of HSP70 family members

lysyl hydroxylase

Keratin 9

Keratin 10

Keratin 1.The identified seven HSP27-interacting proteins by IP coupled with MS could be classified into three functional categories based on their functions:chaperones

cytoskeleton organization

metabolic enzymes.Conclusion:HSP27 in HCC is a related proteins involved in the regulation

by interacting with some peptides

or small molecule protein to control the tumor development.

关键词

热休克蛋白27肝癌免疫共沉淀液相色谱-电喷雾离子阱质谱蛋白质印迹

Keywords

HSP27hepatocellular carcinomaCo-immunoprecipitationLC-ESI-ITMS/MSWestern blot

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