The First Cytoplasmic Loop in the Core Structure of the ABCC1 (Multidrug Resistance Protein 1; MRP1) Transporter Contains Multiple Amino Acids Essential for Its Expression

• Published in: International journal of molecular sciences Vol. 22; no. 18; p. 9710
• Main Authors: Conseil, Gwenaëlle, Cole, Susan P. C.
• Format: Journal Article
• Language: English
• Published: Basel MDPI AG 08-09-2021; MDPI
• Subjects: ABC transporter; Adenosine triphosphate; Alanine; Amino acids; Anions; Binding; Binding sites; cytoplasmic loop; Domains; Efflux; Helices; Ion pairs; MRP1; Multidrug resistance; multidrug resistance protein; Multidrug resistant organisms; Mutagenesis; Mutants; Mutation; Nucleotides; organic anion transport; protein expression; Protein transport; Proteins; Residues
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms22189710

ABCC1 (human multidrug resistance protein 1 (hMRP1)) is an ATP-binding cassette transporter which effluxes xeno- and endobiotic organic anions and confers multidrug resistance through active drug efflux. The 17 transmembrane α-helices of hMRP1 are distributed among three membrane spanning domains (MSD0, 1, 2) with MSD1,2 each followed by a nucleotide binding domain to form the 4-domain core structure. Eight conserved residues in the first cytoplasmic loop (CL4) of MSD1 in the descending α-helix (Gly392, Tyr404, Arg405), the perpendicular coupling helix (Asn412, Arg415, Lys416), and the ascending α-helix (Glu422, Phe434) were targeted for mutagenesis. Mutants with both alanine and same charge substitutions of the coupling helix residues were expressed in HEK cells at wild-type hMRP1 levels and their transport activity was only moderately compromised. In contrast, mutants of the flanking amino acids (G392I, Y404A, R405A/K, E422A/D, and F434Y) were very poorly expressed although Y404F, E422D, and F434A were readily expressed and transport competent. Modeling analyses indicated that Glu422 and Arg615 could form an ion pair that might stabilize transporter expression. However, this was not supported by exchange mutations E422R/R615E which failed to improve hMRP1 levels. Additional structures accompanied by rigorous biochemical validations are needed to better understand the bonding interactions crucial for stable hMRP1 expression.