Solubilization of membrane proteins for two-dimensional gel electrophoresis: identification of sarcoplasmic reticulum membrane proteins
Solubilization of membrane proteins for two-dimensional gel electrophoresis: identification of sarcoplasmic reticulum membrane proteins
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DOI：
10.1016/j.ab.2003.10.024
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发表时间：
2004-02-01
影响因子：
2.9
通讯作者：
Periasamy, M
中科院分区：
文献类型：
--
作者：
Babu, GJ;Wheeler, D;Periasamy, M
用于双向电泳（2DE）的膜蛋白的溶解是非常困难的。在这项研究中，我们报告了使用1，2-二庚酰基-sn-甘油-3-磷酸二酰胆碱（DHPC）作为去污剂，以溶解膜蛋白的2DE。以富含肌浆网（SR）膜蛋白的大鼠心室微粒体组分为模型系统。与磺基甜菜碱或ASB-14相比，DHPC与高浓度尿素的相容性增加了蛋白质的溶解度。肽质量分析有助于识别关键SR膜蛋白，包括SR Ca 2 + ATP酶和其他膜蛋白，这是以前没有报道的2DE。这些结果表明，DHPC是一种更好的溶解膜蛋白的去污剂，可用于生成大多数复杂细胞器（包括SR）的蛋白质组图谱。（C）2003 Elsevier Inc. All rights reserved.
Solubilization of membrane proteins for two-dimensional electrophoresis (2DE) is very difficult. In this study, we report the use of 1,2-diheptanoyl-sn-glycero-3-phosphatdiyl choline (DHPC) as a detergent to solubilize integral membrane proteins for 2DE. Rat ventricular microsomal fractions enriched with sarco(endo)plasmic reticulum (SR) membrane proteins were used as a model system. Compatibility of DHPC with a high concentration of urea increases the solubility of proteins compared with sulphobetaines or ASB-14. Peptide mass analysis assisted in the identification of key SR membrane proteins including SR Ca2+ ATPase and other membrane proteins, which have not previously been reported on 2DE. These results suggest that DHPC is a better detergent for solubilizing membrane proteins and may be useful in generating proteomic maps for most complex organelles including SR. (C) 2003 Elsevier Inc. All rights reserved.