Proteomics analysis for the signal transduction under stimulation of prolactin on Jurkat cells
- VernacularTitle:催乳素在Jurkat细胞中信号传导分子的蛋白质组分析
- Author:
Xiahong LIN
;
Ling LIN
- Publication Type:Journal Article
- Keywords:
Prolactin;
Proteomics;
Hot shock protein;
Nuclear receptor co-repressor;
Galactose-1-phosphate uridyl transferase;
Zinc finger
- From:
Chinese Journal of Immunology
2000;0(08):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:The experiment was designed to apply proteomics to analyze the signal transduction molecules in T lymphocyte strain JurkatD1.1 cells with the stimulation of prolactin.Methods:JurkatD1.1 cells were cultured in the presence of recombinant human prolactin(rhPRL).Phosphated metal affinity chromography(PMAC) and immunoprecipitation(IP) assay were applied to enrich phosphoproteins from the total cell lystates which were resolved by one-dimensional electrophoresis(1 DE) or 2 DE.The different bands and different spots in the gels between control group and rhPRL-treated group were excised,digested,and analyzed by MALDI-TOF mass spectrometry.Results:Three major protein bands were observed specifically in the rhPRL-treated group and other two major protein bands were observed specifically in the control group.The bands were excised and analyzed by MALDI-TOF mass spectrometry.Protein band a in rhPRL-treated group was successfully identified as hot shock protein 90(hsp90).Nine different protein spots which were observed in the 2 DE gels between the two groups were excised and analyzed by MALDI-TOF mass spectrometry.In the rhPRL-treated group the protein spot 4 was identified as nuclear receptor co-repressor 2 variant(NcoR 2 variant);The protein spot 5 was identified as galactose-1-phosphate uridyl transferase;And the protein spot 6 was identified as zinc finger ZIM3.Conclusion:Hot shock protein 90 participates the signal transduction pathways of prolactin.Nuclear receptor co-repressor 2 variant and zinc finger ZIM3 may play a role in the process of prolactin modulating target genes' expression.
