Supplementary Material

Evaluating Fmoc-Amino Acids as Selective Inhibitors of Butyrylcholinesterase

Jeannette Gonzalez, Jennifer Ramirez, and Jason P. Schwans

Page S1: Index of Supplementary Material

Page S2: Supplementary Table 1 Effect of Fmoc-amino acids on BChE activity

Page S3: Supplementary Table 2 Effect of Fmoc-amino acids on AChE activity

Page S4: Supplementary Table 3 IC50 values for inhibition of BChE using Fmoc-amino acids

Page S5: Supplementary Table 4 Calculated molecular van der Waals volumes of Fmoc-amino acids

Page S6: Supplementary Table 5 Effect of amino acids on BChE activity

Page S7: Supplementary Table 6 Effect of amino acids on AChE activity

Page S8: Supplementary Table 7 Effect of Cbz-amino acids on BChE activity

Page S9: Supplementary Table 8 Effect of Boc-amino acids on BChE activity

Page S10: Supplementary Table 9 Inhibition constants (KI values) for inhibition of BChE using Fmoc-amino acids bearing side chain protecting groups

Supplementary Table 1. Effect of Fmoc-amino acids on BChE activity
Amino acid / Relative activity (%) / Amino acid / Relative activity (%)
Fmoc-Ala-O- / 99 ± 1 / Fmoc-Lys-O- / 43 ± 1
Fmoc-Arg-O- / 81 ± 8 / Fmoc-Met-O- / 89 ± 3
Fmoc-Asn-O- / 101 ± 6 / Fmoc-Phe-O- / 58 ± 2
Fmoc-Asp-O- / 100 ± 2 / Fmoc-Pro-O- / 87 ± 6
Fmoc-Gln-O- / 97 ± 4 / Fmoc-Ser-O- / 86 ± 1
Fmoc-Glu-O- / 82 ± 3 / Fmoc-Thr-O- / 96 ± 2
Fmoc-Gly-O- / 100 ± 1 / Fmoc-Trp-O- / 54 ± 1
Fmoc-His-O- / 90 ± 5 / Fmoc-Tyr-O- / 68 ± 3
Fmoc-Ile-O- / 70 ± 3 / Fmoc-Val-O- / 72 ± 2
Fmoc-Leu-O- / 24 ± 7
Supplementary Table 2. Effect of Fmoc-amino acids on AChE activity
Amino acid / Relative activity (%) / Amino acid / Relative activity (%)
Fmoc-Ala-O- / 101 ± 2 / Fmoc-Lys-O- / 94 ± 4
Fmoc-Arg-O- / 100 ± 2 / Fmoc-Met-O- / 100 ± 3
Fmoc-Asn-O- / 104 ± 7 / Fmoc-Phe-O- / 100 ± 6
Fmoc-Asp-O- / 100 ± 2 / Fmoc-Pro-O- / 101 ± 2
Fmoc-Gln-O- / 103 ± 2 / Fmoc-Ser-O- / 104 ± 3
Fmoc-Glu-O- / 99 ± 2 / Fmoc-Thr-O- / 99 ± 3
Fmoc-Gly-O- / 97 ± 4 / Fmoc-Trp-O- / 100 ± 2
Fmoc-His-O- / 98 ± 1 / Fmoc-Tyr-O- / 97 ± 4
Fmoc-Ile-O- / 100 ± 2 / Fmoc-Val-O- / 100 ± 5
Fmoc-Leu-O- / 100 ± 3
Supplementary Table 3. IC50 values for inhibition of BChE using Fmoc-amino acids
Fmoc-amino acid / IC50 (mM)
Fmoc-Leu-O- / 134 ± 15
Fmoc-Lys-O- / 153 ± 14
Fmoc-Ile-O- / 182 ± 19
Fmoc-Val-O- / 322 ± 23
Fmoc-Phe-O- / 320 ± 10
Fmoc-Tyr-O- / 389 ± 24
Fmoc-Trp-O- / 255 ± 27

a van der Waals volumes from Table 3 are reproduced here for completeness

Supplementary Table 4. Calculated van der Waals volumes of Fmoc-amino acidsa
Amino acid / van der Waals volume (Å3) / Amino acid / van der Waals volume (Å3)
Fmoc-Ala-O- / 284.76 / Fmoc-Lys-O- / 348.96
Fmoc-Arg-O- / 368.32 / Fmoc-Met-O- / 337.86
Fmoc-Asn-O- / 319.21 / Fmoc-Phe-O- / 357.37
Fmoc-Asp-O- / 315.68 / Fmoc-Pro-O- / 306.99
Fmoc-Gln-O- / 336.51 / Fmoc-Ser-O- / 293.55
Fmoc-Glu-O- / 332.98 / Fmoc-Thr-O- / 310.85
Fmoc-Gly-O- / 267.47 / Fmoc-Trp-O- / 377.07
Fmoc-His-O- / 341.01 / Fmoc-Tyr-O- / 366.16
Fmoc-Ile-O- / 336.65 / Fmoc-Val-O- / 319.35
Fmoc-Leu-O-OO-O- / 336.65
Supplementary Table 5. Effect of amino acids on BChE activity
Amino acid / Relative activity (%) / Amino acid / Relative activity (%)
H-Ala-O- / 94 ± 4 / H-Lys-O- / 99 ± 1
H-Arg-O- / 94 ± 4 / H-Met-O- / 99 ± 4
H-Asn-O- / 97 ± 2 / H-Phe-O- / 99 ± 5
H-Asp-O- / 97 ± 2 / H-Pro-O- / 99 ± 2
H-Gln-O- / 96 ± 4 / H-Ser-O- / 99 ± 2
H-Glu-O- / 93 ± 3 / H-Thr-O- / 97 ± 2
H-Gly-O- / 99 ± 1 / H-Trp-O- / 100 ± 1
H-His-O- / 96 ± 1 / H-Tyr-O- / 97 ± 3
H-Ile-O- / 100 ± 3 / H-Val-O- / 96 ± 1
H-Leu-O- / 97 ± 2
Supplementary Table 6. Effect of amino acids on AChE activity
Amino acid / Relative activity (%) / Amino acid / Relative activity (%)
H-Ala-O- / 96 ± 2 / H-Lys-O- / 96 ± 3
H-Arg-O- / 99 ± 1 / H-Met-O- / 95 ± 2
H-Asn-O- / 98 ± 4 / H-Phe-O- / 98 ± 1
H-Asp-O- / 98 ± 4 / H-Pro-O- / 98 ± 4
H-Gln-O- / 98 ± 3 / H-Ser-O- / 96 ± 5
H-Glu-O- / 98 ± 7 / H-Thr-O- / 97 ± 1
H-Gly-O- / 96 ± 6 / H-Trp-O- / 95 ± 2
H-His-O- / 99 ± 3 / H-Tyr-O- / 94 ± 3
H-Ile-O- / 100 ± 2 / H-Val-O- / 97 ± 2
H-Leu-O- / 97 ± 5
Supplementary Table 7. Effect of Cbz-amino acids on BChE activity
Boc-amino acid / Relative activity (%)
Cbz-Leu-O- / 78 ± 6
Cbz-Ile-O- / 97 ± 1
Cbz-Val-O- / 100 ± 4
Cbz-Phe-O- / 97 ± 5
Cbz-Tyr-O- / 92 ± 1
Cbz-Trp-O- / 92 ± 1
Cbz-Lys-O- / 87 ± 3
Supplementary Table 8. Effect of Boc-amino acids on BChE activity
Boc-amino acid / Relative activity (%)
Boc-Leu-O- / 99 ± 1
Boc-Ile-O- / 97 ± 1
Boc-Val-O- / 100 ± 1
Boc-Phe-O- / 97 ± 1
Boc-Tyr-O- / 95 ± 2
Boc-Trp-O- / 94 ± 4
Boc-Lys-O- / 97 ± 4
Supplementary Table 9. Inhibition constants (KI values) for inhibition of BChE using Fmoc-amino acids bearing side-chain protecting groups
Fmoc-amino acid / KI (mM)
Fmoc-Trp(Boc)-O- / 23 ± 2
Fmoc-Tyr(OtBu)-O- / 331 ± 19
Fmoc-Lys(Z)-O- / 159 ± 10

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