Evaluation of protein extraction protocols for 2DE in marine ecotoxicoproteomics

	Evaluation of protein extraction protocols for 2DE in marine ecotoxicoproteomics
	Wu, Huifeng1 ; Ji, Chenglong1,2 ; Wei, Lei 1,2; Zhao, Jianmin1
发表期刊	PROTEOMICS
ISSN	1615-9853
	2013-11-01
卷号	13 期号:21 页码:3205-3210
关键词	2de Animal Proteomics Ecotoxicoproteomics Protein Extraction Reproducibility
产权排序	[Wu, Huifeng; Ji, Chenglong; Wei, Lei; Zhao, Jianmin] Chinese Acad Sci, Yantai Inst Coastal Zone Res YIC, Key Lab Coastal Zone Environm Proc,YICCAS, Shandong Prov Key Lab Coastal Zone Environm Proc, Yantai 264003, Peoples R China; [Ji, Chenglong; Wei, Lei] Chinese Acad Sci, Grad Sch, Beijing, Peoples R China
通讯作者	Wu, HF (reprint author), Chinese Acad Sci, Yantai Inst Coastal Zone Res, Yantai 264003, Peoples R China. hfwu@yic.ac.cn
英文摘要	In ecotoxicoproteomics, an accurate and reproducible extraction of proteins is a critical step for 2DE analysis and further protein identification using MS. The criteria for the assessment of protein extraction quality include protein yield, protein spots resolved in a 2DE gel, matched protein spots in replicate gels, reproducibility, and compatibility with MS. In this work, we evaluated three protein extraction systems, straightforward lysis buffer, trichloroacetic acid-acetone, and TRIzol reagent with some modifications, for the protein extraction from three animal species including mussel Mytilus galloprovincialis, flounder Paralichthys olivaceus, and polychaete Nereis diversicolor used in marine ecotoxicology. Our results indicated that these methods could extract significantly different protein profiles. The method using TRIzol reagent resulted in the most matched protein spots resolved in four replicate 2DE gels and highest reproducibilities for the gill of M. galloprovincialis and liver of P. olivaceus. However, a modified trichloroacetic acid-acetone solvent system was best for the whole soft tissue of N. diversicolor. This work provides the fundamental information of the extraction quality of protein extraction protocols from different marine animals, which may facilitate the selection of a suitable protein extraction protocol for ecotoxicoproteomics.; In ecotoxicoproteomics, an accurate and reproducible extraction of proteins is a critical step for 2DE analysis and further protein identification using MS. The criteria for the assessment of protein extraction quality include protein yield, protein spots resolved in a 2DE gel, matched protein spots in replicate gels, reproducibility, and compatibility with MS. In this work, we evaluated three protein extraction systems, straightforward lysis buffer, trichloroacetic acid-acetone, and TRIzol reagent with some modifications, for the protein extraction from three animal species including mussel Mytilus galloprovincialis, flounder Paralichthys olivaceus, and polychaete Nereis diversicolor used in marine ecotoxicology. Our results indicated that these methods could extract significantly different protein profiles. The method using TRIzol reagent resulted in the most matched protein spots resolved in four replicate 2DE gels and highest reproducibilities for the gill of M. galloprovincialis and liver of P. olivaceus. However, a modified trichloroacetic acid-acetone solvent system was best for the whole soft tissue of N. diversicolor. This work provides the fundamental information of the extraction quality of protein extraction protocols from different marine animals, which may facilitate the selection of a suitable protein extraction protocol for ecotoxicoproteomics.
文章类型	Article
资助机构	NSFC [21237004]; Natural Science Foundation of Shandong Province [2013JQF01012]; Key Deployment Program of Chinese Academy of Sciences [KZZD-EW-14-03]
收录类别	SCI
语种	英语
关键词[WOS]	HIGH-FAT DIET ; MYTILUS-GALLOPROVINCIALIS ; PROTEOMIC ANALYSIS ; STRESS
研究领域[WOS]	Biochemistry & Molecular Biology
WOS记录号	WOS:000330251700014
引用统计	被引频次：25[WOS] [WOS记录] [WOS相关记录]
文献类型	期刊论文
条目标识符	http://ir.yic.ac.cn/handle/133337/6937
专题	中国科学院海岸带环境过程与生态修复重点实验室海岸带生物学与生物资源利用重点实验室_海岸带生物学与生物资源保护实验室
作者单位	1.Chinese Acad Sci, Yantai Inst Coastal Zone Res YIC, Key Lab Coastal Zone Environm Proc,YICCAS, Shandong Prov Key Lab Coastal Zone Environm Proc, Yantai 264003, Peoples R China 2.Chinese Acad Sci, Grad Sch, Beijing, Peoples R China
推荐引用方式 GB/T 7714	Wu, Huifeng,Ji, Chenglong,Wei, Lei,et al. Evaluation of protein extraction protocols for 2DE in marine ecotoxicoproteomics[J]. PROTEOMICS,2013,13(21):3205-3210.
APA	Wu, Huifeng,Ji, Chenglong,Wei, Lei,&Zhao, Jianmin.(2013).Evaluation of protein extraction protocols for 2DE in marine ecotoxicoproteomics.PROTEOMICS,13(21),3205-3210.
MLA	Wu, Huifeng,et al."Evaluation of protein extraction protocols for 2DE in marine ecotoxicoproteomics".PROTEOMICS 13.21(2013):3205-3210.

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