Figure 1.

Validation of the interaction between CtsD and the A_2AR C-terminal tail in yeast.A, transformation of Yeast 2H Gold strain with pGBKT7, pGBKT7-A_2AR^284–410 (bait), pGADT7 and pGADT7-CtsD^321–410 (prey) vectors in a pairwise combination, according to the numbered yeast line. The yeast cells were grown on SD/Leu⁻ Trp⁻ or SD/Leu⁻ Trp⁻ Ade⁻ His⁻ (in the presence of Aureobasidin and X-α-Gal) selective medium. pGBKT7-p53 and pGADT7-SV40 small T antigen were used as positive controls and pGBKT7-p53 and pGADT7-Lamin expressing vectors were used as negative controls. B, presentation of the different maturation forms of CtsD. Characterization of A_2AR^284–410 interactor clones that encode the different ORFs of the CtsD. The amino acid numbers of pre–pro-CtsD protein (P18242) is based on UniProtKB/Swiss-Prot databases. S: N-terminal secretion signal peptide (20 amino acids [AA]); P: propeptide (44 AA); C: cleavage site between the light and heavy chain of CtsD; arrows indicate the processing sites; D: catalytic Asp residues (D97 and D295) are depicted by black triangles; N: N-linked glycosylation sites at Asn residues (N134 and N261) are depicted by empty triangles (26). A_2AR, adenosine A_2A receptor; CtsD, cathepsin D.