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. Author manuscript; available in PMC: 2010 Oct 15.
Published in final edited form as: Biochim Biophys Acta. 2007 Nov 29;1778(2):454–465. doi: 10.1016/j.bbamem.2007.11.010

Fig. 2. Substitution of S685 in Walker A motif of NBD1 with an amino acid residue that prevents formation of hydrogen-bond with the D792 in Walker B motif significantly affects the protein folding.

Fig. 2

A. Sequence alignment of the Walker A and B motifs in NBD1 of MRP1. The highlighted letters indicate that the serine residue at 685 was substituted with the amino acid residues indicated at that position. The definition of S685A means that the S685 in Walker A motif was substituted with an alanine residue while the Walker B motif was un-mutated. B. Expression of wild-type and variant mutants of human MRP1 proteins in BHK cells at 37 °C. Cells were lysed with 2% SDS and loaded to a 7% polyacrylamide gel. The amounts of proteins loaded in the gel are indicated at the bottom of the gel. Molecular weight markers (kDa) are indicated on the left. MRP1 and im MRP1 on the right indicate the complex-glycosylated mature MRP1 and the core-glycosylated immature MRP1 proteins that were detected in western blot by employing the monoclonal antibody 42.4 against NBD1 [20].