Proteins in the insulin-secreting cell line MIN6 bind the imidazoline compound BL11282.
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 approximately 50 kDa differential band upon SDS-PAGE and a weaker 100 kDa band. The former was sensitive to competitive removal by preincubation of the fraction with BL11282, then highlighting the approximately 100 kDa band. Masspectrometric analysis revealed the approximately 50 kDa band to be EF1A and the approximately 100 kDa band to be glucose regulated P94, both of interest in insulin synthesis and secretion.
Shafqat
Jawed
J
Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden.
Ishrat
Moin
M
Jägerbrink
Theres
T
Sillard
Rannar
R
Mäeorg
Uno
U
Efendic
Suad
S
Berggren
Per-Olof
PO
Zaitsev
Sergei V
SV
Jörnvall
Hans
H
D000595:Amino Acid Sequence; D000818:Animals; D002458:Cell Fractionation; D002460:Cell Line; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D007004:Hypoglycemic Agents / Q000378:metabolism* / Q000494:pharmacology; D007093:Imidazoles / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D050417:Insulin-Secreting Cells / Q000187:drug effects / Q000378:metabolism*; D013058:Mass Spectrometry; D051379:Mice; D008969:Molecular Sequence Data; D020648:Peptide Elongation Factor 1 / Q000032:analysis / Q000378:metabolism*; D011485:Protein Binding; D020349:Surface Plasmon Resonance
C423844:BL 11282; D007004:Hypoglycemic Agents; D007093:Imidazoles; D020648:Peptide Elongation Factor 1