Preliminary Function
Study of NAG7 Using
Two-dimensional Electrophoresis and Mass Spectrometry
TAN Chen, LI Jiang, XIE Yi, XIANG Qiu, WANG Jie-Ru, LIANG
Song-Ping1*, LI Gui-Yuan*
( Cancer Research Institute, Xiang Ya Medical College, Central South
University,Changsha 410078, China；1College of Life Science, Hunan
Normal University, Changsha 410081, China )
Abstract In search
of mechanisms of function of NAG7 gene, a powerful new tool for the
unambiguous characterization of gel-separated proteins is accomplished by the
combination of mass spectrometry and sequence database searching. NAG7,
a novel putative tumor suppressor gene, located on 3p25.3, was introduced into
HNE1 cells by liposome transfection. We used two dimensional polyacrylamide gel
electrophoresis (2-D PAGE) to identify proteins that were overexpressed in NAG7
transfected cells. After staining and image analysis, spots of interest were
isolated and subjected to mass spectrometry (MS). We found nine proteins, which
up-regulated in NAG7 transfected cells and be identified by MS. These
proteins included growth arrest specific protein, DNA binding protein, c-myc
promoter-binding protein and caspase 6 etc., which involved in cell cycling,
transcription regulation, and apoptosis. NAG7 may exert the function
by up-regulating the expression of these proteins.
Key words NAG7 gene； cell transfection； two-dimensional gel
electrophoresis； matrix-assisted laser
desorption/ionization mass spectrometry
*Corresponding author： LI Gui-Yuan： Tel, 86-731-4805446； Fax, 86-731-4805383； e-mail, [email protected]； LIANG Song-Ping： Tel, 86-731-8872556； Fax,86-731-8861304；
e-mail,[email protected]