ABCMdb

PMID: 12084728

Milewski MI, Mickle JE, Forrest JK, Stanton BA, Cutting GR
Aggregation of misfolded proteins can be a selective process dependent upon peptide composition.
J Biol Chem. 2002 Sep 13;277(37):34462-70. Epub 2002 Jun 25., 2002-09-13 [PubMed]

Sentences

No. Mutations Sentence Comment

85 ABCC7 p.Val1397Glu The only reported CF-causing mutation in the ag region was a substitution of valine at codon 1397 by glutamic acid (V1397E) (29). Login to comment 88 ABCC7 p.Thr1396Ala The replacement of Thr-1396 with alanine had no effect on aggregation of the GFP-tagged protein (Fig. 1D). Login to comment 90 ABCC7 p.His1402Ala ABCC7 p.Arg1403Ala Only 25% (52/208) of cells expressing GFP fusion protein bearing single H1402A substitution and 27% (41/153) of R1403A mutants showed intracellular protein accumulation, whereas the corresponding number for the wild type sequence was 91% (132/145). Login to comment 93 ABCC7 p.His1402Ala ABCC7 p.Arg1403Ala Cells expressing the double mutant H1402A,R1403A showed no aggregation at all (0/108). Login to comment 97 ABCC7 p.His1402Ala ABCC7 p.Arg1403Ala However, the introduction of the ⌬ag deletion or the H1402A,R1403A double mutation significantly decreased the amount of the fusion protein in the insoluble fraction to approximately 31 and 36%, respectively. Login to comment 108 ABCC7 p.His1402Ala ABCC7 p.Arg1403Ala ABCC7 p.Val1397Glu ABCC7 p.Thr1396Ala B-E, the effect of different mutations within the ag region, including ⌬ag (B), V1397E (C), T1396A (D), and double mutation H1402A,R1403A (E) on the aggregation of C terminus of CFTR fused to GFP (shown in green) in transiently transfected human airway epithelial (IB3-1) cells. Login to comment 120 ABCC7 p.His1402Ala ABCC7 p.Arg1403Ala Additionally, the double mutation H1402A,R1403A prevented aggregation of HA-tagged CFTR C terminus (Fig. 2E). Login to comment 125 ABCC7 p.Ser1455* To examine whether this C-terminal region contributed to the unusual accumulation pattern of HA-CFTR 1370-1480, we deleted the last 26 amino acids in this construct by introducing the naturally occurring CFTR mutation S1455X (33), associated with abnormal localization of the full-length protein (31). Login to comment 146 ABCC7 p.His1402Ala ABCC7 p.Arg1403Ala E, lack of aggregation of HA-CFTR 1370-1480 construct carrying the H1402A and R1403A mutations. Login to comment 160 ABCC7 p.His1402Ala ABCC7 p.Arg1403Ala Similarly, the non-aggregating GFP-CFTR 1370-1480 H1402A,R1403A construct was included in giant aggregates formed by HA-CFTR 1370-1480 (Fig. 4D). Login to comment 195 ABCC7 p.His1402Ala ABCC7 p.His1402Ala ABCC7 p.Arg1403Ala ABCC7 p.Arg1403Ala C, colocalization of the aggregating GFP-CFTR 1370-1480 construct (green) and the non-aggregating HA-CFTR 1370-1480 H1402A,R1403A construct (red) in giant aggregates. Login to comment 196 ABCC7 p.His1402Ala ABCC7 p.Arg1403Ala D, co-localization of the non-aggregating GFP-CFTR 1370-1480 H1402A,R1403A construct (green) with the aggregating HA-CFTR 1370-1480 construct (red) in giant aggregates. Login to comment