RÉSUMÉ
TRIM28, known as a kind of macro-molecules, consists of multiple domains and belongs to one of human tripartite motif-containing proteins families. This superfamily contains more than 60 proteins in total and is characterized by four conservative domains, also referred to the RBCC domain, which is a RING (really interesting new gene) finger, two B-boxes (type 1 and type 2) and a leucine zipper coiled-coil region. TRIM28 plays its pivotal roles in transcriptional co-activation or co-repression by interacting directly with transcription factors, within which is a Kruppel-associated box domain (KRAB domain). Moreover, it is critical during the development of embryos, the differentiation of cells and the regulation of cancers.
RÉSUMÉ
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.