Differential Protein Expression Induced by Transient
Transfection of Metallothionein-3 Gene in SH-SY5Y Neuroblastoma Cell Line
ZHOU Bo, YANG Wei, JI Jian-Guo*, RU Bing-Gen*
(
Proteome Group, National Laboratory of Protein Engineering, College of Life
Sciences, Peking University, Beijing 100871, China )
Abstract Metallothionein-3
(MT-3), also known as growth inhibitory factor (GIF), is predominantly
expressed in central nervous system (CNS). It belongs to the family of
metallothionein (MT) but has several unique properties that are not shared by
other family members such as MT-1/MT-2. In the past few years, MT-3 had been
postulated to be a multipurpose protein which could play important
neuromodulatory and neuroprotective roles in CNS besides the common roles of
MTs. However, the primary function of MT-3 and the mechanism underlying its
multiple functions were not elucidated so far. In present study, human
neuroblastoma cell line SH-SY5Y was employed to study the overall cellular
protein changes induced by transient transfection of MT-3 gene, based on
comparative proteome analysis. Averagely about 750 spots were visualized by
Coomassie staining in one 2D gel, in which 17 proteins were shown to display
significant and reproducible changes by semiquantitative analysis with
ImageMaster 2D Elite software. Among them, 12 proteins were up-regulated while
other 5 proteins were down-regulated. Using matrix-assisted laser
desorption/ionization time-of-flight mass spectrometry, 10 proteins were
further identified to be zinc finger protein, glutamate transporter, and
enhancer protein, etc., which were involved in several important pathways
regulating the functions of central nervous system. The results showed that
MT-3 might exert its unique functions by regulating the expression of these
proteins.
Key words metallothionein-3;
cell transfection; 2-DE; MALDI-TOF-MS
*Corresponding author：
RU Bing-Gen: Tel, 86-10-62751842; Fax,
86-10-62751842; e-mail, [email protected];
JI Jian-Guo: Tel, 86-10-62753115; e-mail,
[email protected]