Proteins in the insulin-secreting cell line MIN6 bind the imidazoline compound BL11282

Shafqat, Jawed; Ishrat, Moin; Jägerbrink, Theres; Sillard, Rannar; Mäeorg, Uno; Efendic, Suad; Berggren, Per-Olof; Zaitsev, Sergei V.; Jörnvall, Hans

doi:10.1016/j.febslet.2008.04.007

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Volume 582, Issue 11 , Pages 1613-1617, 14 May 2008

Proteins in the insulin-secreting cell line MIN6 bind the imidazoline compound BL11282

Edited by Robert Barouki

Jawed Shafqat
- Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden
Moin Ishrat
- Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden
Theres Jägerbrink
- Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden
Rannar Sillard
- Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden
Uno Mäeorg
- Institute of Chemistry, University of Tartu, Tartu, Estonia
Suad Efendic
- The Rolf Luft Research Center for Diabetes and Endocrinology, Karolinska Institutet, Stockholm, Sweden
Per-Olof Berggren
- The Rolf Luft Research Center for Diabetes and Endocrinology, Karolinska Institutet, Stockholm, Sweden
Sergei V. Zaitsev
- The Rolf Luft Research Center for Diabetes and Endocrinology, Karolinska Institutet, Stockholm, Sweden
- Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow, Russian Federation
Hans Jörnvall
- Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden
- Corresponding author. Fax: +46 8 33 74 62.

Received 16 March 2008; accepted 7 April 2008. published online 15 April 2008.

Abstract

The imidazoline BL11282 stimulates insulin release and alters islet proteomes. Subcellular fractions of MIN6 cells showed that the membrane fraction exhibited binding to BL11282 on a Biacore chip and to BL11282-labelled magnetic beads. Bound material extracted from the beads showed a ∼50kDa differential band upon SDS–PAGE and a weaker 100kDa band. The former was sensitive to competitive removal by preincubation of the fraction with BL11282, then highlighting the ∼100kDa band. Masspectrometric analysis revealed the ∼50kDa band to be EF1A and the ∼100kDa band to be glucose regulated P94, both of interest in insulin synthesis and secretion.