Evaluation of the SELDI-TOF MS technique for protein profiling of pancreatic islets exposed to glucose and oleate.

Abstract:

:The aim of the study was to evaluate the SELDI-TOF MS technique for pancreatic islet research. Mouse islets were cultured at low or high glucose levels in the absence or presence of oleate and characterized by measuring insulin secretion and oxygen tension. Subsequently, the islets were protein profiled. Up to 200 different peaks could be detected in a single experiment with the majority of peaks corresponding to proteins with masses below 30 kDa. By combining different protein arrays, the number of detected peaks could be increased further. The optimal binding of islet proteins was achieved using the anionic exchange array and phosphate buffer (pH 6) when the binding of insulin was low, which allowed other less abundant proteins to be captured. When islets from different culture conditions were profiled and analyzed, in total 25 proteins were found to be oleate/glucose-regulated. An oleate-regulated protein was chosen for identification work, which was conducted by passive elution from SDS-PAGE gels and subsequent in-gel trypsin digestion and MALDI-TOF MS. The protein was identified as peptidyl-prolyl isomerase B (PPI-B). In conclusion, the study demonstrates that SELDI-technique can be used not only to obtain islet protein patterns but is also helpful in the subsequent identification of differentially expressed proteins.

journal_name

Proteomics

journal_title

Proteomics

authors

Ortsäter H,Sundsten T,Lin JM,Bergsten P

doi

10.1002/pmic.200601019

subject

Has Abstract

pub_date

2007-09-01 00:00:00

pages

3105-15

issue

17

eissn

1615-9853

issn

1615-9861

journal_volume

7

pub_type

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