ABCMdb

PMID: 26545497

Cuviello F, Tellgren-Roth A, Lara P, Ruud Selin F, Monne M, Bisaccia F, Nilsson I, Ostuni A
Membrane insertion and topology of the amino-terminal domain TMD0 of multidrug-resistance associated protein 6 (MRP6).
FEBS Lett. 2015 Dec 21;589(24 Pt B):3921-8. doi: 10.1016/j.febslet.2015.10.030. Epub 2015 Nov 3., [PubMed]

Sentences

No. Mutations Sentence Comment

39 ABCC6 p.Glu125Lys R60GY deletion was introduced in constructs where TM1 is in LepH2 and in LepH3 and E125K was introduced in constructs where TM3 and TM4 are in both Lep model proteins. Login to comment 60 ABCC6 p.Ser245Thr To exclude the possibility that the lack of glycosylation in the C-terminal tail was due to the presence of a serine instead of a threonine in the glycosylation site, one of them was mutated into threonine (S245T TMD0-L0). Login to comment 128 ABCC6 p.Glu125Lys ABCC6 p.Glu125Lys TMD0 TMs Predicted DG (kcal/mol) Membrane insertion efficiency (%) Measured DG (kcal/mol) Positive charges H2 H3 H2 H3 N-term C-term TM1 Q16-K72 0.707 82 72 0.87 0.56 1 4 TM2 H57-P102 0.922 86 87 1.08 1.03 4 1 TM3 K93-Q131 1.628 54 72 0.09 0.56 1 3 TM4 H123-R166 1.029 92 31 1.45 0.47 3 1 TM5 N150-T206 0.020 93 86 1.52 1.05 1 0 TM1-DR60GY 0.707 85 71 1.03 0.52 1 3 TM3-E125K 1.628 48 80 0.05 0.82 1 4 TM4-E125K 1.029 92 38 1.43 0.30 4 1 F. Cuviello et al. / FEBS Letters xxx (2015) xxx-xxx 5 displayed cleaved fragments and it contains a SignalP predicted cleavage site between A187 and D188 of the original TMD0-L0 sequence. Login to comment 147 ABCC6 p.Glu125Lys The deletion of R60GY [43] in the C-terminal flanking region of TM1 was introduced in the Lep constructs with TM1 and the E125K mutation [44] in the loop between TM3 and TM4 was introduced in Lep constructs with TM3 and TM4. Login to comment 165 ABCC6 p.Glu125Lys The R60GY deletion that reduce the number of positively charged residues in the C-terminal cytoplasmic flanking region of TM1 and the E125K substitution that adds a positive charge in the short cytoplasmic loop between TM3 and TM4 both cause PXE. Login to comment 168 ABCC6 p.Glu125Lys The E125K mutation increases the positive charge difference between the cytosolic and lumenal sides of TM3 and TM4 from 3 against 1 to 4 against 1 and therefore in theory this substitution increases their natural topological preferences. Login to comment