ABCMdb

ABCC4 p.Arg815Ala

None has been submitted yet.

Publications

PMID: 23766510 [PubMed] Cheepala SB et al: "Crucial role for phylogenetically conserved cytoplasmic loop 3 in ABCC4 protein expression."

No. Sentence Comment

207 Mutation R815A in Human ABCC4 Cytoplasmic Region 3 (CL3) Destabilizes the Protein-Because CLs interact with NBDs in CFTR (26), we hypothesized that a mutation in an amino acid of CL3 in the "coupling helix" (a CL region near the NBD) might disrupt NBD and CL interdomain interactions producing an alteration in stability. Login to comment
209 To investigate this we mutated Arg-815 to Ala. Login to comment
210 The R815A mutation resulted in reduced ABCC4 expression and loss of mature glycosylated band c (Fig. 6, B and C). Login to comment
211 The reduction in mature ABCC4 is consistent with the prediction that CL3 Arg-815 interacts with the NBD to ensure proper folding of the protein To test if reduced temperature enhanced expression of R815A, NIH3T3 cells were transiently transfected with ABCC4, T796M-ABCC4, or R815A-ABCC4 and then incubated at 28 &#b0;C. Login to comment
212 The amount of R815A protein increased, which paralleled an increase in mature glycosylated protein band c (Fig. 6, B and C). Login to comment
257 The increased formation of the mature c-form of ABCC4 R815A at the subphysiological temperature of 28 &#b0;C is consistent with the proposal that CL3/NBD1 interactions are necessary for proper folding of ABCC4. Login to comment
267 A, representation of interdomain interaction between R815A of CL3 region and Asp-522 of NBD1 domain. Login to comment
268 B, immunoblot of whole cell lysates (100 òe;g of protein per lane) prepared from NIH3T3 cells transfected with human ABCC4 WT and ABCC4 mutant (R815A) and incubated at 37 or 28 &#b0;C for 24 h after 24 h of transfection. Login to comment
278 Likewise, we show that the predicted contact between CL3 Arg-815 and NBD1 Asp-522 when disrupted by an R815A substitution reduces the amount of mature ABCC4. Login to comment
206 Mutation R815A in Human ABCC4 Cytoplasmic Region 3 (CL3) Destabilizes the Protein-Because CLs interact with NBDs in CFTR (26), we hypothesized that a mutation in an amino acid of CL3 in the "coupling helix" (a CL region near the NBD) might disrupt NBD and CL interdomain interactions producing an alteration in stability. Login to comment
208 To investigate this we mutated Arg-815 to Ala. Login to comment
256 The increased formation of the mature c-form of ABCC4 R815A at the subphysiological temperature of 28 &#b0;C is consistent with the proposal that CL3/NBD1 interactions are necessary for proper folding of ABCC4. Login to comment
266 A, representation of interdomain interaction between R815A of CL3 region and Asp-522 of NBD1 domain. Login to comment
277 Likewise, we show that the predicted contact between CL3 Arg-815 and NBD1 Asp-522 when disrupted by an R815A substitution reduces the amount of mature ABCC4. Login to comment