Proteome Analysis of Interleukin-1β–Induced Changes in Protein Expression in Rat Islets of Langerhans
- P. Mose Larsen1,
- S.J. Fey1,
- M.R. Larsen2,
- A. Nawrocki1,
- H.U. Andersen3,
- H. Kähler3,
- C. Heilmann3,
- M.C. Voss3,
- P. Roepstorff2,
- F. Pociot3,
- A.E. Karlsen3 and
- J. Nerup3
- 1Center for Proteome Analysis and the
- 2Institute for Biochemistry and Molecular Biology, University of Southern Denmark, Odense
- 3Steno Diabetes Center, Gentofte, Denmark
Abstract
The intracellular molecular events involved in the β-cell death process are complex but poorly understood. Cytokines, e.g., interleukin (IL)-1β, may play a crucial role in inducing this process. Protein synthesis is necessary for the deleterious effect of IL-1, and induction of both protective and deleterious proteins has been described. To characterize the rather complex pattern of islet protein expression in rat islets in response to IL-1, we have attempted to identify proteins of altered expression level after IL-1 exposure by 2D gel electrophoresis and mass spectrometry. Of 105 significantly changed (i.e., up- or downregulated or de novo–induced) protein spots, we obtained positive protein identification for 60 protein spots. The 60 identifications corresponded to 57 different proteins. Of these, 10 proteins were present in two to four spots, suggesting that posttranslatory modifications had occurred. In addition, 11 spots contained more than one protein. The proteins could be classified according to their function into the following groups: 1) energy transduction; 2) glycolytic pathway; 3) protein synthesis, chaperones, and protein folding; and 4) signal transduction, regulation, differentiation, and apoptosis. In conclusion, valuable information about the molecular mechanisms involved in cytokine-mediated β-cell destruction was obtained by this approach.
Footnotes
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Address correspondence and reprint requests to Dr. Peter Mose Larsen, Center for Proteome Analysis, Forskerparken Fyn, Odense, Denmark. E-mail: nerup{at}pres.dk.
Received for publication 6 September 2000 and accepted in revised form 5 February 2001.
BSA, bovine serum albumin; DMEM, Dulbecco’s modified Eagle’s medium; HBSS, Hanks’ balanced salt solution; HSP, heat shock protein; IEF, isoelectric focusing; IL, interleukin; iNOS, inducible nitric oxide synthase; MALDI, matrix-assisted laser desorption/ionization; MS, mass spectrometry; NEPHGE, nonequilibrium pH-gradient electrophoresis; NHS, normal human serum; NO, nitric oxide; pI, isoelectric point; SOD, super oxide dismutase; TCA, trichloroacetic acid.














