Artificial Super-Receptor Interface Mimic and Exceed Protein Functionality

Abstract

Active sites in proteins account for a small proportion but are crucial for their enhanced binding affinity and specificity, making related biomimetic structures a research hotspot. However, current structures greatly depended on rigid inorganic frameworks for high-certainty assembly, which introduced interfering inorganic groups and interactions not present in proteins. To address this, we utilized organic crystal rigidity to achieve high-certainty assembly conformations. Thus periodic active sites at crystal interface can be precisely assembled by pure organic units. Our three-step strategy for designing artificial super-receptors includes: (1) Learning active site model from proteins; (2) Imitating active sites in crystal cell unit; (3) Exceeding natural performance with periodic active sites at the crystal interface. Practically, by mimicking the human dopamine transporter (amphetamine drug receptor), our artificial super-receptor acted as super-sensor. It achieved a limit of detection down to 480 pM, 64,580 times lower than the natural receptor. It also showed revolutionary broad-spectrum specificity for amphetamine drugs, including chiral methamphetamine, ecstasy, and even potential novel amphetamine derivative illicit drugs, allowing active preventing detection for the drug abuse problem. The customized designing strategy was also validated by high dopamine sensitivity (2.8 nM) and selectivity of d-DAT inspired PySO₃H artificial super-receptor. Such strategy can be further extended to other functional proteins for various super-performance, from sensor, catalyst, medicine, agriculture to therapeutic applications, etc.

Keywords

artificial super-receptor / broad-spectrum specific sensor / chiral drug recognition / micro crystal interface / protein active site

References

[1] M. Cianci, Introduction to Proteins: Structure, Function, and Motion, 2nd Edition, in Crystallography Reviews (CRC Press, 2021), 47–50.

[2] G. E. Schulz and R. H. Schirmer, Principles of Protein Structure (Springer Science & Business Media, 2013).

[3] A. Kessel and N. Ben-Tal, Introduction to Proteins: Structure, Function, and Motion (Chapman and Hall/CRC, 2018).

[4] L. Gao, J. Zhuang, L. Nie, et al., “Intrinsic Peroxidase-Like Activity of Ferromagnetic Nanoparticles,” Nature Nanotechnology 2 (2007): 577–583.

[5] J. Sheng, Y. Wu, H. Ding, et al., “Multienzyme-Like Nanozymes: Regulation, Rational Design, and Application,” Advanced Materials 36 (2023): 2211210.

[6] R. Zhang, X. Yan, and K. Fan, “Nanozymes Inspired by Natural Enzymes,” Accounts of Materials Research Journal 2 (2021): 534–547.

[7] H. Wang, K. Wan, and X. Shi, “Recent Advances in Nanozyme Research,” Advanced Materials 31 (2018): 1805368.

[8] X. Cai, L. Jiao, H. Yan, et al., “Nanozyme-Involved Biomimetic Cascade Catalysis for Biomedical Applications,” Materials Today 44 (2021): 211–228.

[9] J. Zhang and J. Liu, “Nanozyme-Based Luminescence Detection,” Luminescence 35 (2020): 1185–1194.

[10] X. Wang, L. Qin, M. Zhou, Z. Lou, and H. Wei, “Nanozyme Sensor Arrays for Detecting Versatile Analytes From Small Molecules to Proteins and Cells,” Analytical Chemistry 90 (2018): 11696–11702.

[11] Y. Huang, J. Ren, and X. Qu, “Nanozymes: Classification, Catalytic Mechanisms, Activity Regulation, and Applications,” Chemical Reviews 119 (2019): 4357–4412.

[12] D. Jiang, D. Ni, Z. T. Rosenkrans, P. Huang, X. Yan, and W. Cai, “Nanozyme: New Horizons for Responsive Biomedical Applications,” Chemical Society Reviews 48 (2019): 3683–3704.

[13] M. Liang and X. Yan, “Nanozymes: From New Concepts, Mechanisms, and Standards to Applications,” Accounts of Chemical Research 52 (2019): 2190–2200.

[14] I. W. Hamley, “Protein Assemblies: Nature-Inspired and Designed Nanostructures,” Biomacromolecules 20 (2019): 1829–1848.

[15] A. Nekrasov, Y. Kozmin, S. Kozyrev, R. Ziganshin, A. de Brevern, and A. Anashkina, “Hierarchical Structure of Protein Sequence,” International Journal of Molecular Sciences 22 (2021): 8339.

[16] M. Heim, L. Romer, and T. Scheibel, “Hierarchical Structures Made of Proteins. The Complex Architecture of Spider Webs and Their Constituent Silk Proteins,” Chemical Society Reviews 39 (2010): 156–164.

[17] X. Liu, X. Xu, D. Hilger, et al., “Structural Insights Into the Process of GPCR-G Protein Complex Formation,” Cell 177 (2019): 1243–1251.

[18] J. W. Bryson, S. F. Betz, H. S. Lu, et al., “Protein Design: A Hierarchic Approach,” Science 270 (1995): 935–941.

[19] A. S. Ali, E. Shakhnovich, and M. Karplus, “How Does a Protein Fold?,” Nature 369 (1994): 248–251.

[20] C. M. Dobson, “Protein Folding and Misfolding,” Nature 426 (2003): 884–890.

[21] X. Feng, X. Wang, C. Redshaw, and B. Z. Tang, “Aggregation Behaviour of Pyrene-Based Luminescent Materials, From Molecular Design and Optical Properties to Application,” Chemical Society Reviews 52 (2023): 6715–6753.

[22] F. P. Kinik, A. Ortega-Guerrero, D. Ongari, C. P. Ireland, and B. Smit, “Pyrene-Based Metal Organic Frameworks: From Synthesis to Applications,” Chemical Society Reviews 50 (2021): 3143–3177.

[23] Y. Q. Zhao, L. Yu, L. Zhang, H. Liu, Y. Zhou, and J. S. Kim, “Activated Aggregation-Induced Emission Therapeutics Agents for Triggering Regulated Cell Death,” Aggregate 5 (2024): e503.

[24] A. Sharma, P. Verwilst, M. Li, et al., “Theranostic Fluorescent Probes,” Chemical Reviews 124 (2024): 2699–2804.

[25] L. Shi, Y. H. Liu, K. Li, et al., “An AIE-Based Probe for Rapid and Ultrasensitive Imaging of Plasma Membranes in Biosystems,” Angewandte Chemie International Edition 59 (2019): 9962–9966.

[26] W. Xu, H. Liu, F. Mei, et al., “Supramolecule-Originated Emission: A Room-Temperature Phosphorescence 2D Ionic H-Bond Network From Nonemissive Aliphatic Derivatives,” ACS Applied Materials & Interfaces 13 (2021): 61528–61535.

[27] W. Xu, Y. Fu, H. Liu, Q. He, H. Cao, and J. Cheng, “Attogram “Meth” Detection Enabled by Selective Organic Crystal Disaggregation via Directed Crystal Level Interactions,” Analytical Chemistry 90 (2018): 1402–1407.

[28] H. Liu, Y. Fu, W. Xu, et al., “Microcrystal Induced Emission Enhancement of a Small Molecule Probe and Its Use for Highly Efficient Detection of 2,4,6-Trinitrophenol in Water,” Science China Chemistry 61 (2018): 857–862.

[29] Y. Zhuang, P. Xu, C. Mao, et al., “Structural Insights Into the Human D1 and D2 Dopamine Receptor Signaling Complexes,” Cell 184 (2021): 931–942.

[30] Y. Zhuang, B. Krumm, H. Zhang, et al., “Mechanism of Dopamine Binding and Allosteric Modulation of the Human D1 Dopamine Receptor,” Cell Research 31 (2021): 593–596.

[31] J. Frank, J. Zhu, P. Penczek, et al., “A Model of Protein Synthesis Based on Cryo-Electron Microscopy of the E. coli Ribosome,” Nature 376 (1995): 441–444.

[32] X. Teng, S. Chen, Y. Nie, et al., “Ligand Recognition and Biased Agonism of the D1 Dopamine Receptor,” Nature Communications 13 (2022): 3186.

[33] S. Kondo, T. Takahashi, N. Yamagata, et al., “Neurochemical Organization of the Drosophila Brain Visualized by Endogenously Tagged Neurotransmitter Receptors,” Cell reports 30 (2020): 284–297.

[34] J. Y. Hansen, G. Shafiei, R. D. Markello, et al., “Mapping Neurotransmitter Systems to the Structural and Functional Organization of the Human Neocortex,” Nature Neuroscience 25 (2022): 1569–1581.

[35] K. H. Wang, A. Penmatsa, and E. Gouaux, “Neurotransmitter and Psychostimulant Recognition by the Dopamine Transporter,” Nature 521 (2015): 322–327.

[36] D. Ding, K. Li, B. Liu, and B. Z. Tang, “Bioprobes Based on AIE Fluorogens,” Accounts of Chemical Research 46 (2013): 2441–2453.

[37] J. Han, J. You, X. Li, P. Duan, and M. Liu, “Full-Color Tunable Circularly Polarized Luminescent Nanoassemblies of Achiral AIEgens in Confined Chiral Nanotubes,” Advanced Materials 29 (2017): 1606503.

RIGHTS & PERMISSIONS

2026 The Author(s). Aggregate published by SCUT, AIEI, and John Wiley & Sons Australia, Ltd.