DICP OpenIR
Subject Area分析化学
Effect of alkyl chain length on reversed phase stationary phase for separation of integral membrane protein digests
Zhao Q(赵群); Sun LL(孙良亮); Wu Q(吴琪); Liang Z(梁振); Zhang LH(张丽华); Zhang YK(张玉奎)
Source Publication26th International Symposium on Microscale Bioseparations
Conference Name26th International Symposium on Microscale Bioseparations
Conference Date2011-5-1
2011
Conference Place圣地亚哥
Pages75-0
Publisher待补充
Publication Place待补充
Cooperation Status墙报
Department1810
Funding OrganizationCASSS
AbstractIntegral membrane proteins (IMPs) play a critical role in regulating various cellular processes, including intercellular communication, vesicle trafficking, ion transport, protein translocation/integration, and the propagation of signaling cascades. However, the IMP analysis presents great challenge due to the hydrophobic nature. Recently, much attention has been paid to the solubilization of IMPs by chaotropes, detergents, organic solvents, and organic acid. Besides, the separation of hydrophobic peptide also has significant impact on the identification of IMPs. However, the in-depth study on this aspect has rarely been reported. In our recent work, membrane proteins from rat hippocampus were first solubilized by ionic liquids based protocol [1]. Then, to decrease the complexity of membrane protein digests, the prefractionation of peptides was performed during desalting. Finally, the effect of alkyl chain length of stationary phase on the separation of IMP digests was studied by μRPLC-ESI-MS/MS. In duplicate runs, in total, 536 and 703 peptides, corresponding to 301 and 398 proteins (FDR <5% at peptide level), were identified with protein digests separated by C18 and C8, respectively. For hydrophobic peptides and IMPs, the identified number obtained by C8 column was improved by 29.3% and 20.3%, respectively, compared with that obtained with C18 stationary phase. All these results indicate that C8 reverse-phase stationary phase is preferred for the large scale membrane proteome study; Integral membrane proteins (IMPs) play a critical role in regulating various cellular processes, including intercellular communication, vesicle trafficking, ion transport, protein translocation/integration, and the propagation of signaling cascades. However, the IMP analysis presents great challenge due to the hydrophobic nature. Recently, much attention has been paid to the solubilization of IMPs by chaotropes, detergents, organic solvents, and organic acid. Besides, the separation of hydrophobic peptide also has significant impact on the identification of IMPs. However, the in-depth study on this aspect has rarely been reported. In our recent work, membrane proteins from rat hippocampus were first solubilized by ionic liquids based protocol [1]. Then, to decrease the complexity of membrane protein digests, the prefractionation of peptides was performed during desalting. Finally, the effect of alkyl chain length of stationary phase on the separation of IMP digests was studied by μRPLC-ESI-MS/MS. In duplicate runs, in total, 536 and 703 peptides, corresponding to 301 and 398 proteins (FDR <5% at peptide level), were identified with protein digests separated by C18 and C8, respectively. For hydrophobic peptides and IMPs, the identified number obtained by C8 column was improved by 29.3% and 20.3%, respectively, compared with that obtained with C18 stationary phase. All these results indicate that C8 reverse-phase stationary phase is preferred for the large scale membrane proteome study
Language英语
Document Type会议论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/116024
Collection中国科学院大连化学物理研究所
Corresponding AuthorZhang LH(张丽华)
Recommended Citation
GB/T 7714
Zhao Q,Sun LL,Wu Q,et al. Effect of alkyl chain length on reversed phase stationary phase for separation of integral membrane protein digests[C]. 待补充:待补充,2011:75-0.
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