Dried polyacrylamide gel absorption: A method for efficient elimination of the interferences from SDS-solubilized protein samples in mass spectrometry-based proteome analysis

2010 ◽  
Vol 31 (23-24) ◽  
pp. 3816-3822 ◽  
Author(s):  
Jian Zhou ◽  
Jianglin Li ◽  
Jianjun Li ◽  
Ping Chen ◽  
Xianchun Wang ◽  
...  
2008 ◽  
Vol 71 (2) ◽  
pp. 160-167 ◽  
Author(s):  
Kelan Zhang ◽  
Krzysztof Wrzesinski ◽  
Stephen J. Fey ◽  
Peter Mose Larsen ◽  
Xumin Zhang ◽  
...  

Author(s):  
Fatemeh Nasri ◽  
Maryam Zare ◽  
Mehrnoosh Doroudchi ◽  
Behrouz Gharesi-Fard

Background: Polycystic ovary syndrome (PCOS) is the most frequent endocrine disorder affecting 6–7% of premenopausal women. Recent studies revealed that the immune system especially CD4+ T helper cells are important in the context PCOS. Proteome analysis of CD4+ T lymphocytes can provide valuable information regarding the biology of these cells in the context of PCOS. Objective: To investigate immune dysregulation in CD4+ T lymphocytes at the protein level in the context of PCOS using two-dimensional gel electrophoresis (2DE) and mass spectrometry (MS). Methods: In the present study, we applied two-dimensional gel electrophoresis / mass spectrometry to identify proteins differentially expressed by peripheral blood CD4+ T cells in ten PCOS women compared with ten healthy women. Western blot technique was used to confirm the identified proteins. Results: Despite the overall proteome similarities, there were significant differences in the expression of seven spots between two groups (P <0.05). Three proteins, namely phosphatidylethanolamine-binding protein 1, proteasome activator complex subunit 1 and triosephosphate isomerase 1 were successfully identified by Mass technique and confirmed by western blot. All characterized proteins were over-expressed in CD4+ T cells from patients compared to CD4+ T cells from controls (P <0.05). In-silico analysis suggested that the over-expressed proteins interact with other proteins involved in cellular metabolism especially glycolysis and ferroptosis pathway. Conclusion: These findings suggest that metabolic adjustments in CD4+ T lymphocytes, which is in favor of increased glycolysis and Th2 differentiation are important in the context of PCOS.


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