Proteome analysis of rat pancreas induced by pancreatectomy.
The previous study demonstrated that the streptozotocin (STZ)-induced diabetic mice can be cured by injecting the regenerating pancreatic extract (RPE) of the partially pancreatectomized Wistar-Kyoto rats. In this study, to characterize the complex pattern of protein expression in RPE, the proteins of altered expression level after the pancreatectomy were identified by 2-dimensional electrophoresis (2-DE) and mass spectrometry. Of 76 significantly up- or down-regulated protein spots, 61 were identified by MALDI-TOF/MS. Moreover, the whole RPE was fractionated into 4 groups using an anion-exchange chromatography and each fraction's cell proliferating activity was measured by MTT assay. Compared to the normal pancreatic extract, fraction 3 and 4 of RPE showed the maximal cell proliferating activity. On 2-DE of 3 and 4 fractions, a total of 10 spots, which are differentially expressed after the pancreatectomy, were identified by MS/MS. Of these identified proteins, Reg III which might be functionally associated with well known regenerating factor (Reg I) was found. Taken together, our results demonstrated that the differential protein expression associated with pancreas regeneration could be sought by 2-DE and mass spectroscopy and suggested that the pre-fractionation method combined with in vitro cell proliferation assay is effectively used to pinpoint the active components for pancreas regeneration.
Shin
Jun-Seop
JS
Graduate School of Life Sciences and Biotechnology, Korea University, Seoul 136-701, South Korea.
Lee
Jae-Jeong
JJ
Lee
Eun-Ju
EJ
Kim
Yeon-Hee
YH
Chae
Kwon-Seok
KS
Kim
Chan-Wha
CW
D000818:Animals; D049109:Cell Proliferation; D015180:Electrophoresis, Gel, Two-Dimensional; D010179:Pancreas / Q000378:metabolism*; D010180:Pancreatectomy; D011506:Proteins / Q000032:analysis* / Q000378:metabolism*; D020543:Proteome / Q000032:analysis* / Q000378:metabolism*; D051381:Rats; D011921:Rats, Inbred WKY; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D011506:Proteins; D020543:Proteome