Dr. Jae-Won Yang has been working on comparative and functional proteomics based on mass spectrometry. With respect to functional proteomics of ion channels, he has identified in vivo phosphorylation sites in voltage-gated potassium Kv1 channels and numerous interacting proteins in mouse, rat and human brain tissues using mass spectrometry. Using the relatively quantitative “Stable Isotope Labeling by Amino acids in Cell Culture (SILAC)” technique, trafficking dependent-phosphorylation was characterized for surface expression of Kv1.2 channels. Only a few research groups do perform ion channel studies regarding in vivo post-translational modification as the hydrophobicity of these integral membrane proteins renders purification and identification difficult. He has applied this research method for Kv7 channels and expanded for dopamine transporter (DAT) and serotonin transporter (SERT) to study their modulation by phosphorylation.
All the infrastructure, equipment and instruments required for the research such as confocal and fluorescence microscopes and mass spectrometers (QTRAP and MALDI-TOF/TOF) are available at the Center of Physiology and Pharmacology.
Techniques available in his research group are as follow:
Mass Spectrometry – Identification of protein and post-translational modification (phosphorylation and glycosylation),
Stable Isotope Labeling with Amino acids in Cell culture (SILAC) quantification,
Two-dimensional gel electrophoresis.
SDS-PAGE, Western blot (1D and 2D), Immunoprecipitation,
Affinity purification, Phospho-specific antibody work, Immunofluorescence, cell surface biotinylation, cell cultures.
Site-directed mutagenesis, DNA/RNA purification.
Foster JD, Yang JW, Moritz AE, Challasivakanaka S, Smith MA, Holy M, Wilebski K, Sitte HH, Vaughan RA. Dopamine transporter phosphorylation site threonine 53 regulates substrate reuptake and amphetamine-stimulated efflux. J Biol Chem. 2012 Jun 21. [Epub ahead of print]
Vacher H, Yang JW, Cerda O, Autillo-Touati A, Dargent B, Trimmer JS (2011) CDK-mediated phosphorylation of Kvß2 auxiliary subunit regulates Kv1 channel axonal targeting. J Cell Biol 192:813-824.
Park KS, Yang JW, Seikel EL, Trimmer JS (2008) Potassium channel phosphorylation in excitable cells: Providing dynamic functional variability to a diverse family of ion channels. Physiology 23:49-57
Yang JW, Vacher H, Park KS, Clark E, Trimmer JS (2007) Trafficking-dependent phosphorylation of Kv1.2 regulates voltage-gated potassium channel cell surface expression. Proc Natl Acad Sci USA 104 (50):20055-20060
Yang JW, Afjehi-Sadat L, Gelpi E, Kunze M, Hoger H, Fleckner J, Berger J, Lubec G (2006) Proteome profiling in the rat Harderian gland. J Proteome Res 5(7):1751-1762