Figure 1
From: Insulin receptor membrane retention by a traceable chimeric mutant
Covalent modification of IR-B at the plasma membrane. A. Tag position and insulin binding scheme according to proposed model [33, 34]. L1, Large leucine rich domain 1; CR, cysteine rich domain; L2, large leucine rich domain 2; FnIII domains 1, 2 and 3; ID, inter-domain; TM, transmembrane domain; TK, tyrosine kinase; CT, C-terminus. Inserted tag shows: in grey, residues added for cloning reasons; in red, Ser recognized by ACP-S; in blue, conserved flanking amino acids for enzyme recognition [30]. B. Labeling strategy. C-E. HeLa cells expressing Mut-GFP (C-D) or Mut (E) were labeled with 0.2 μM ACP-S and 1 μM fluorescent CoA and imaged by confocal (Mut-GFP) or wide field microscopy (Mut). Scale bars: 10 μm. D. Manders map. F. Cells transiently expressing wild type IR-B or the mutant were assayed by Western blot 48 h after transfection using anti-IR-β-subunit or anti-ERK1/2. Quantification was performed by densitometry. Results are expressed as the mean ± s.e.m. (*: p≤0.05; n≥3).