From 2001 to 2005, my research focus was the role of protein turn-over in cardiac protection. Specifically, proteasome is the major protease in the heart. When the heart is under stress, the production of damaged proteins drastically increases. Such proteins need to be removed in a timely fashion; otherwise they will induce toxic effects. In these circumstances, proteasome regulation is one of the main mechanisms to counteract the stress. I have developed methodology to purified proteasome from murine heart tissue and analyzed the composition of this protein complex. Furthermore, I explored the role of phosphorylation in tuning the functionality of this protein complex. Proteasome regulating kinase/phosphatase has been identified and the targets of such modifications reported. Besides my own project, I also actively participated in other on-going projects investigating cardiac protection signaling network, including mitochondria related apoptosis and kinase signaling cascade. The overall goal of the research is to illustrate the molecular mechanism of cardiac protection and find suitable targets for possible clinical interventions.
Over the years, I have mastered multiple biological techniques, including Fast Protein Liquid Chromatography (FPLC: ion-exchange chromatography, gel-filtration chromatography, hydrophobic-interaction chromatography, reverse-phase chromatography and affinity chromatography), western blotting, Immunoprecipitation, Enzyme-Linked Immunosorbent Assay (ELISA), 1D SDS PAGE, 2D electrophoresis, Native gel electrophoresis, blue native gel electrophoresis, zymogen electrophoresis, DNA agarose electrophoresis, polymerase chain reaction, recombinant protein expression and purification (both bacteria and insect cells), mammalian cell culture, kinase activity assay and protease activity assay. Furthermore, I have personally pursued and very familiar with immunohistochemistry, confocal microscopy, electron microscopy, MALDI-TOF mass spectrometry and HPLC coupled tandem mass spectrometry. My strength is combined biology and proteomics background.