ABCMdb

PMID: 23316740

Sampson HM, Lam H, Chen PC, Zhang D, Mottillo C, Mirza M, Qasim K, Shrier A, Shyng SL, Hanrahan JW, Thomas DY
Compounds that correct F508del-CFTR trafficking can also correct other protein trafficking diseases: an in vitro study using cell lines.
Orphanet J Rare Dis. 2013 Jan 14;8:11. doi: 10.1186/1750-1172-8-11., [PubMed]

Sentences

No. Mutations Sentence Comment

24 ABCC8 p.Val187Asp ABCC8 p.Ala116Pro We investigated mutants from a diverse set of well-studied protein trafficking diseases including the nephrogenic diabetes insipidus mutations V206D and L292P in the arginine-vasopressin receptor 2 (AVPR2, also known as V2R) [24,25], the LQTS2 mutations G601S and F805C in the human ether-a-go-go-related gene (KCNH2, also known as hERG) [26,27], and finally the persistent hyperinsulinemic hypoglycemia of infancy (PHHI, also known as congenital hyperinsulinism) mutations A116P and V187D in the sulfonylurea receptor 1 (ABCC8, also known as SUR1) [10,28,29], a component of the KATP channel. Login to comment 34 ABCC8 p.Val187Asp ABCC8 p.Ala116Pro FLAG-tagged hamster SUR1 WT, A116P and V187D mutants and rat Kir6.2 plasmids have been described previously [32]. Login to comment 120 ABCC8 p.Val187Asp ABCC8 p.Ala116Pro 6 F508del-CFTR correctors correct the trafficking of SUR1 mutants V187D and A116P. Login to comment 121 ABCC8 p.Val187Asp ABCC8 p.Ala116Pro Representative immunoblots are shown for SUR1 V187D (A-C) and SUR1 A116P (D-F). Login to comment 122 ABCC8 p.Val187Asp Representative immunoblot for cells expressing SUR1 V187D treated with decreasing concentrations of ouabain (ouab), KM57, and RDR1 (A), KM60, ABT-888, and latonduine (latond) (B), glafenine and carbamazepine (carbam) (C). Login to comment 123 ABCC8 p.Ala116Pro Representative immunoblot for cells expressing SUR1 A116P treated with 10% glycerol (glycerol), decreasing concentrations of ouabain and glafenine (D), carbamazepine (carbam), KM57, and KM60 (E), latonduine, RDR1, ABT-888 (F). Login to comment 125 ABCC8 p.Val187Asp ABCC8 p.Ala116Pro Lanes where Kir6.2 and SUR1 V187D or SUR1 A116P are expressed are indicated by a line above the corresponding lanes. Login to comment 131 ABCC8 p.Val187Asp ABCC8 p.Ala116Pro We next tested the SUR1 mutants V187D and A116P for correction with F508del-CFTR correctors. Login to comment 134 ABCC8 p.Ala116Pro In contrast to cells expressing wild type SUR1 protein, those expressing the SUR1 A116P mutant showed only low levels of channel activity due to loss of surface expression after treatment with the DMSO control. Login to comment 135 ABCC8 p.Ala116Pro Exposing A116P-expressing cells to the reversible sulfonylurea drug tolbutamide for > 24 h followed by a washout 2 h prior to the assay led to almost complete recovery of channel activity, as reported previously [32]. Login to comment 136 ABCC8 p.Ala116Pro Among the correctors Figure 7 The F508del-CFTR corrector RDR1 improves the function of the SUR1 A116P mutant. Login to comment 137 ABCC8 p.Ala116Pro (A) Representative 86 Rb+ efflux profiles from COS cells transiently transfected with SUR1 A116P and wt Kir6.2 and treated with 0.1% DMSO, 300bc;M tolbutamide (a reversible sulfonylurea) or 10bc;M RDR1 as described in the METHODS. Login to comment 154 ABCC8 p.Ala116Pro that enhanced the processing of A116P, KM57, KM60 and RDR1 were tested for their effects on functional recovery of the mutant channel. Login to comment 158 ABCC8 p.Val187Asp ABCC8 p.Ala116Pro Interestingly, sulfonylureas correct the trafficking of A116P and V187D mutants by binding to sites outside the affected domain in SUR1, and possibly also with a weak affinity site in Kir6.2 [36]. Login to comment 167 ABCC8 p.Val187Asp ABCC8 p.Ala116Pro Table 1 F508del-CFTR corrector compounds show distinct profiles of correction for other ER-retained proteins Corrector CFTR F508del hERG G601S hERG F805C SUR1 A116P SUR1 V187D V2R L292P V2R V206D VRT-325 + + - - ND - ND Glycerol + ND ND + ND ND +/- 29&#b0;C ++ + + + + ++ ++ KM60 + + - + + - + KM57 +/- ++ - +/- - - +/- ABT-888 + - ND + + - + Glafenine + + - + - - - RDR1 + - - + + - - Ouabain + +/-* +* + + + + Carbamazepine + - ND - +/- - ND Latonduine + +/- - + ++ +/- + Astemizole ND + - ND ND ND ND Glibenclamide ND ND ND ++ ++ ND ND A qualitative assessment of correction as determined by glycosylation status in immunoblotting is shown for each mutation following treatment with a corrector compound, where "-" indicates no correction observed, "+/-" indicates slight correction, "+" and "++" indicate more and best correction observed, respectively, and "ND" indicates not determined. Login to comment