An integrated approach to proteome analysis: identification of proteins associated with cardiac hypertrophy.
An integrated approach to proteome analysis: identification of proteins associated with cardiac hypertrophy.
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蛋白质组分析的综合方法：鉴定与心脏肥大相关的蛋白质。
DOI：
10.1006/abio.1998.2566
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发表时间：
1998
影响因子：
2.9
通讯作者：
J. Stults
中科院分区：
文献类型：
--
作者：
D. Arnott;K. O'Connell;K. King;J. Stults
心肌细胞肥大是心脏对超负荷的主要反应，是心力衰竭和死亡的独立预测因子。不同的细胞表型与由不同原因引起的肥大相关。这些表型已经被其他人通过分析基因转录模式在分子水平上描述。另一种方法是通过二维聚丙烯酰胺凝胶电泳分析大规模蛋白质表达模式（蛋白质组）。实现这一目标需要能够严格分析复杂的2D凝胶图像，有效地消化单个凝胶分离的蛋白质（特别是那些以低水平表达的蛋白质），并以高灵敏度分析所得肽以进行快速数据库搜索。我们致力于改进技术和实验方法来应对这些挑战，以便有效地研究心肌肥大的细胞培养模型。分析了来自有或没有苯肾上腺素诱导的肥大的心肌细胞的多个样品的细胞裂解物的2D凝胶图案，并定位了具有统计学显著性的丰度变化的点。十一个这样的斑点，确定使用改进的程序，在凝胶消化银染蛋白质和高灵敏度质谱。将低水平的十二烷基硫酸钠掺入消化缓冲液中改善了肽回收率。结合基质辅助激光解吸质谱的初始测量和毛细管液相色谱-离子阱质谱的肽序列测定产生了有效的蛋白质鉴定。2D凝胶图像分析和常规鉴定的蛋白质存在于凝胶中的subpicomole水平的整合代表了蛋白质组研究的一般模型与蛋白质表达模式的基因组序列。
Hypertrophy of cardiac myocytes is a primary response of the heart to overload, and is an independent predictor of heart failure and death. Distinct cellular phenotypes are associated with hypertrophy resulting from different causes. These phenotypes have been described by others at the molecular level by analysis of gene transcription patterns. An alternative approach is the analysis of large-scale protein expression patterns (the proteome) by two-dimensional polyacrylamide gel electrophoresis. Realization of this goal requires the ability to rigorously analyze complex 2D gel images, efficiently digest individual gel isolated proteins (especially those expressed at low levels), and analyze the resulting peptides with high sensitivity for rapid database searches. We have undertaken to improve the technology and experimental approaches to these challenges in order to effectively study a cell culture model for cardiac hypertrophy. The 2D gel patterns for cell lysates from multiple samples of cardiac myocytes with or without phenylephrine-induced hypertrophy were analyzed and spots which changed in abundance with statistical significance were located. Eleven such spots were identified using improved procedures for in-gel digestion of silver-stained proteins and high-sensitivity mass spectrometry. The incorporation of low levels of sodium dodecyl sulfate into the digestion buffer improved peptide recovery. The combination of matrix-assisted laser desorption mass spectrometry for initial measurements and capillary liquid chromatography-ion trap mass spectrometry for peptide sequence determination yielded efficient protein identification. The integration of 2D gel image analysis and routine identification of proteins present in gels at the subpicomole level represents a general model for proteome studies relating genomic sequence with protein expression patterns.
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DOI：
10.1073/pnas.84.20.6970
发表时间：
1987-10-01
影响因子：
11.1
作者：
AEBERSOLD, RH;LEAVITT, J;KENT, SBH
通讯作者：
KENT, SBH
快速、高可信度蛋白质鉴定的策略。
影响因子：
7.4
作者：
Qin,J;Fenyö,D;Zhao,Y;Hall,WW;Chao,DM;Wilson,CJ;Young,RA;Chait,BT
通讯作者：
Chait,BT
影响因子：
7.4
作者：
DAVIS, MT;STAHL, DC;LEE, TD
通讯作者：
LEE, TD
用于二维凝胶定量分析的 QUEST 系统。
DOI：
--
发表时间：
1989
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Garrels,JI
通讯作者：
Garrels,JI
影响因子：
37.8
作者：
CALDERONE, A;TAKAHASHI, N;COLUCCI, WS
通讯作者：
COLUCCI, WS