Identification of growth factor-regulated proteins using 2D electrophoresis and mass spectrometry.
Proteomic technology has recently emerged as a powerful tool for detecting both qualitative and quantitative changes of proteins that occur upon activation of complex signaling pathways. In the present study, comparison of the protein profile of platelet-derived growth factor (PDGF), epidermal growth factor (EGF) and nerve growth factor (NGF)-stimulated and unstimulated cells with two-dimensional electrophoresis followed by mass spectrometric analysis led to the identification of a number of proteins, several of which had not been previously shown to be regulated by receptor-tyrosine kinases. Using subcellular fractionation, our approach was able to identify not only changes due to altered gene transcription, but also due to intracellular protein translocation or modification. One of the proteins that was identified among other PDGF-regulated molecules was prohibitin, a potential tumour suppressor previously implicated in cell cycle regulation and protection of mitochondrial proteins from degradation. Further analysis confirmed that mitochondria-associated prohibitin translocates to an insoluble perinuclear compartment. This study demonstrates the utility of proteomic strategies in identifying potential growth factor-regulated effectors.
Saridaki
Aggeliki
A
Biomedical Sciences Research Center "Alexander Fleming", Laboratory of Protein Chemistry, Vari, Greece.
Panayotou
George
G
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D015180:Electrophoresis, Gel, Two-Dimensional; D004815:Epidermal Growth Factor / Q000494:pharmacology; D005786:Gene Expression Regulation; D006133:Growth Substances / Q000494:pharmacology*; D006801:Humans; D008969:Molecular Sequence Data; D020932:Nerve Growth Factor / Q000494:pharmacology; D010455:Peptides / Q000737:chemistry; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology; D000091402:Prohibitins; D020543:Proteome / Q000235:genetics / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D051381:Rats; D012097:Repressor Proteins / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization
D006133:Growth Substances; D010455:Peptides; D010982:Platelet-Derived Growth Factor; D000091402:Prohibitins; D020543:Proteome; D012097:Repressor Proteins; D004815:Epidermal Growth Factor; D020932:Nerve Growth Factor