Table 3. Linker sequences utilized for creating concatenated α4β2 nAChR constructs
Linked setFull nameC-term.Linker sequenceMature N term.C tailLinkerN term. (to L anchor)Total
Dimeric constructs
β-6-αβ2-6-α4APSSKEG(AGS)6RAHAEE2321650
β-9a-αβ2-9a-α4APSSK(AGS)3AHAEE239638
β-6a-αβ2-6a-α4APSSK(AGS)2AHAEE236635
β-3a-αβ2-3a-α4APSSKAGSAHAEE233632
β-0a-αβ2-0a-α4HSAPSGSAHAEE212629
β-(-3a)-αβ2-(-3a)-α4HSDHSGSAHAEE182626
Tetrameric and pentameric constructs
β1-21a-α2-β21-21a-α42-APSSK(AGS)7AHAEE2321650
β1-3a-α2-β21-3a-α42-APSSKAGSAHAEE233632
23--α42-33a-β23-LAGMI(AGS)5LGS(AGS)5TDTEE833647
23--α42-33a-α43-LAGMI(AGS)5LGS(AGS)5AHAEE833647
34--β23-18a-α44-APSSK(AGS)2AGT(AGS)3AHAEE2318647
34--α43-33a-β24-LAGMI(AGS)7ATG(AGS)3TDTEE833647
45-α44-33a-α45LAGMI(AGS)4ATG(AGS)6AHAEE833647
45-β24-20a-α45APSSKTG(AGS)6AHAEE2320649

The five most proximal C-terminal (C-term.) amino acids and the five first amino acids from the predicted mature N terminus (N term.) are presented for α4 and β2 subunits along with the specific linker sequences introduced during concatenation. C-terminal tail lengths were obtained from NCBI (SwissProt accession nos. P43681 for α4 and P17787 for β2), whereas N-terminal lengths were derived from the definition of an α-helical hydrophobic leucine anchor as described in the Results. For tetrameric and pentameric constructs, numbers indicate the construct position of each subunit (e.g., α2: α4 subunit in the second construct position).