Spatial Structure and Activity of Synthetic Fragments of Lynx1 and of Nicotinic Receptor Loop C Models
Abstract
:1. Introduction
2. Materials and Methods
2.1. Synthesis of Linear and Disulfide-Stabilized Fragments of Human Lynx1, of the Lymnaea Stagnalis AChBP Loop C and of the Combinatorial Peptide HAP
2.2. Activity of Synthetic Fragments of Lynx1 and of Nicotinic Receptor Loop C Models in Radioligand Assay
2.3. CD Analysis of the Synthetic Peptides
2.4. NMR Spectroscopy
3. Results
3.1. Synthesis of Linear and Disulfide-Stabilized Fragments of Human Lynx1 Loop II, of the L. stagnalis AChBP Loop C and of the Combinatorial Peptide HAP
3.2. Activity of Synthetic Fragments of Lynx1 and of Nicotinic Receptor Loop C Models in Radioligand Assay
3.3. CD Analysis of the Synthetic Peptides
3.4. NMR Analysis of Synthetic Lynx1 Fragments
3.5. NMR Analysis of Synthetic Analogs of the L. stagnalis AChBP Loop C
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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№ | Protein Fragment | Sequence | Theoretical Mass/Measured Mass, Da | Abbreviation |
---|---|---|---|---|
(1) | Human Lynx1 fragment 29–43 (Loop II). | TTRTYYTPTRMKVSK | 1833.0/1832.9 | Ly29–43 |
(2) | Human Lynx1 fragment 29–43 with cysteines at N- and C- termini, linear form. | CTTRTYYTPTRMKVSKC | 2039.0/2039.0 | Ly29–43C,C |
(3) | Human Lynx1 fragment 29–43 with cysteines at N- and C-termini, cyclized form. | 2037.0/2037.1 | Ly29–43C-C | |
(4) | L. stagnalis AChBP fragment 202–214 (Loop C), linear form. | SVTYSCCPEAYED | 1466.5/1466.4 | Ls202–214C,C |
(5) | L. stagnalis AChBP fragment 202–214 (Loop C), cyclized form. | 1464.5/1464.6 | Ls202–214C-C | |
(6) | L. stagnalis AChBP fragment 202–214 with cysteines 207–208 mutated to serines and cysteines added at N- and C- termini, linear form. | CSVTYSSSPEAYEDC | 1640.6/1640.6 | mLs202–214C,C |
(7) | L. stagnalis AChBP fragment 202–214 with cysteines 207–208 mutated to serines and cysteines added at N- and C- termini, cyclized form. | 1638.6/1638.6 | mLs202–214C-C | |
(8) | Combinatorial high-affinity peptide HAP. | WRYYESSLLPYPD | 1688.8/1688.7 | HAP |
(9) | Combinatorial high-affinity peptide HAP with cysteines at N- and C- termini, linear form. | CWRYYESSLLPYPDC | 1894.8/1894.8 | HAPC,C |
(10) | Combinatorial high-affinity peptide HAP with cysteines at N- and C- termini, cyclized form. | 1892.8/1892.9 | HAPC-C |
Peptide | α-Helix, % | β-Structure, % | β-Turn, % | Random, % | NRMSD 1 |
---|---|---|---|---|---|
(1) | 5.2 | 31.5 | 22.6 | 40.7 | 0.02 |
(2) | 5.1 | 32.6 | 22.5 | 39.8 | 0.02 |
(3) | 4.1 | 38.5 | 22.5 | 34.9 | 0.03 |
(4) | 5.9 | 32.4 | 23.4 | 38.2 | 0.02 |
(5) | 6.1 | 30.9 | 23.7 | 39.2 | 0.02 |
(6) | 6.4 | 36.1 | 23.0 | 34.5 | 0.03 |
(7) | 5.9 | 33.6 | 22.4 | 38.0 | 0.02 |
(8) | 3.3 | 39.5 | 22.6 | 34.6 | 0.03 |
(9) | 3.9 | 38.5 | 22.3 | 35.3 | 0.03 |
(10) | 3.7 | 39.9 | 21.7 | 34.7 | 0.05 |
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Mineev, K.S.; Kryukova, E.V.; Kasheverov, I.E.; Egorova, N.S.; Zhmak, M.N.; Ivanov, I.A.; Senko, D.A.; Feofanov, A.V.; Ignatova, A.A.; Arseniev, A.S.; et al. Spatial Structure and Activity of Synthetic Fragments of Lynx1 and of Nicotinic Receptor Loop C Models. Biomolecules 2021, 11, 1. https://doi.org/10.3390/biom11010001
Mineev KS, Kryukova EV, Kasheverov IE, Egorova NS, Zhmak MN, Ivanov IA, Senko DA, Feofanov AV, Ignatova AA, Arseniev AS, et al. Spatial Structure and Activity of Synthetic Fragments of Lynx1 and of Nicotinic Receptor Loop C Models. Biomolecules. 2021; 11(1):1. https://doi.org/10.3390/biom11010001
Chicago/Turabian StyleMineev, Konstantin S., Elena V. Kryukova, Igor E. Kasheverov, Natalia S. Egorova, Maxim N. Zhmak, Igor A. Ivanov, Dmitry A. Senko, Alexey V. Feofanov, Anastasia A. Ignatova, Alexander S. Arseniev, and et al. 2021. "Spatial Structure and Activity of Synthetic Fragments of Lynx1 and of Nicotinic Receptor Loop C Models" Biomolecules 11, no. 1: 1. https://doi.org/10.3390/biom11010001