Unraveling hot spots in binding interfaces: progress and challenges
Abstract
Protein interface hot spots, as revealed by alanine scanning mutagenesis, continue to stimulate interest in the biophysical basis of molecular recognition. Although these regions apparently constitute fertile grounds for intermolecular interactions, no general algorithm has yet been developed that can predict hot spots based solely on their shape or composition. The discovery of structural plasticity in hot spot regions indicates that dynamic simulation techniques may be essential for achieving a predictive understanding of binding interface energetics. Future progress will depend as much on the application of new computational approaches for dissecting protein interfaces as on expanding our empirical databank of mutagenic substitutions and their effects. Despite our current theoretical shortcomings, recent methodological advances provide efficient experimental means of probing hot spots and enable immediate applications for hot spots in drug discovery.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
Binding energy is not evenly distributed among interfacial residues.
“It has been verified that the binding energy is not evenly distributed among the interfacial residues.”
From iSEE: Interface structure, evolution, and energy-based machine learning predictor of binding affinity changes upon mutations (2019) · cites this paper as [17]
Some interfacial residues (hot-spots) contribute most to binding affinity.
“Instead, some residues (hot‐spots) contribute most to the binding affinity.16, 17, 18”
From iSEE: Interface structure, evolution, and energy-based machine learning predictor of binding affinity changes upon mutations (2019) · cites this paper as [17]
Cites
Principles of protein-protein interactions
Jones, S., & Thornton, J. M. (1996) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.93.1.13 · Cited by 2,684 · see citing works
Anatomy of hot spots in protein interfaces
Bogan, A. A., & Thorn, K. S. (1998) · Journal of Molecular Biology · doi:10.1006/jmbi.1998.1843 · Cited by 1,988 · see citing works
A Hot Spot of Binding Energy in a Hormone-Receptor Interface
Clackson, T., & Wells, J. A. (1995) · Science · doi:10.1126/science.7529940 · Cited by 1,987 · see citing works
The atomic structure of protein-protein recognition sites 1 1Edited by A. R. Fersht
Conte, L. L., Chothia, C., & Janin, J. (1999) · Journal of Molecular Biology · doi:10.1006/jmbi.1998.2439 · Cited by 1,967 · see citing works
Crystal Structure of a Neutralizing Human IgG Against HIV-1: A Template for Vaccine Design
Saphire, E., Parren, P., Pantophlet, R., Zwick, M., Morris, G., Rudd, P., Dwek, R., Stanfield, R., Burton, D., & Wilson, I. A. G. (2001) · Science · doi:10.1126/science.1061692 · Cited by 985 · see citing works
Massova, I., & Kollman, P. A. (1999) · Journal of the American Chemical Society · doi:10.1021/ja990935j · Cited by 778 · see citing works
Folding and binding cascades: Dynamic landscapes and population shifts
Kumar, S., Ma, B., Tsai, C., Sinha, N., & Nussinov, R. (2000) · Protein Science · doi:10.1110/ps.9.1.10 · Cited by 595 · see citing works
Wang, W., Donini, O., Reyes, C. M., & Kollman, P. A. (2001) · Annual Review of Biophysics and Biomolecular Structure · doi:10.1146/annurev.biophys.30.1.211 · Cited by 549 · see citing works
Understanding protein folding via free-energy surfaces from theory and experiment
Dinner, A. R., Šali, A., Smith, L. J., Dobson, C. M., & Karplus, M. (2000) · Trends in Biochemical Sciences · doi:10.1016/s0968-0004(00)01610-8 · Cited by 496 · see citing works
Site-directed ligand discovery
Erlanson, D. A., Braisted, A. C., Raphael, D. R., Randal, M., Stroud, R. M., Gordon, E. M., & Wells, J. A. (2000) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.97.17.9367 · Cited by 466 · see citing works
Dragic, T., Trkola, A., Thompson, D. A. D., Cormier, E. G., Kajumo, F., Maxwell, E., Lin, S. W., Ying, W., Smith, S. O., Sakmar, T. P., et al. (2000) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.090576697 · Cited by 427 · see citing works
Thorn, K. S., & Bogan, A. A. (2001) · Bioinformatics · doi:10.1093/bioinformatics/17.3.284 · Cited by 403 · see citing works
Rapid mapping of protein functional epitopes by combinatorial alanine scanning
Weiss, G. A., Watanabe, C. K., Zhong, A. W., Goddard, A. D., & Sidhu, S. S. (2000) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.160252097 · Cited by 352 · see citing works
Dissection of a Nuclear Localization Signal
Hodel, M. R., Corbett, A. H., & Hodel, A. E. (2001) · Journal of Biological Chemistry · doi:10.1074/jbc.m008522200 · Cited by 338 · see citing works
BiGGER: A new (soft) docking algorithm for predicting protein interactions
Palma, P. N., Krippahl, L., Wampler, J. E., & Moura, J. J. G. (2000) · Proteins Structure Function and Bioinformatics · doi:10.1002/(sici)1097-0134(20000601)39:4 · Cited by 292 · see citing works
Wet and dry interfaces: the role of solvent in protein–protein and protein–DNA recognition
Janin, J. (1999) · Structure · doi:10.1016/s0969-2126(00)88333-1 · Cited by 224 · see citing works
Piehler, J., Roisman, L. C., & Schreiber, G. (2000) · Journal of Biological Chemistry · doi:10.1074/jbc.m006854200 · Cited by 152 · see citing works
Wang, W., Lim, W. A., Jakalian, A., Wang, J., Wang, J., Luo, R., Bayly, C. I., & Kollman, P. A. (2001) · Journal of the American Chemical Society · doi:10.1021/ja003164o · Cited by 134 · see citing works
Energetic Determinants of Internal Motif Recognition by PDZ Domains
Harris, B. Z., Hillier, B. J., & Lim, W. A. (2001) · Biochemistry · doi:10.1021/bi010142l · Cited by 130 · see citing works
Protein functional epitopes: hot spots, dynamics and combinatorial libraries
Ma, B., Wolfson, H. J., & Nussinov, R. (2001) · Current Opinion in Structural Biology · doi:10.1016/s0959-440x(00)00216-5 · Cited by 111 · see citing works
Scott, J. K., Huang, S., Gangadhar, B. P., Samoriski, G. M., Clapp, P., Gross, R. A., Taussig, R., & Smrcka, A. V. (2001) · The EMBO Journal · doi:10.1093/emboj/20.4.767 · Cited by 103 · see citing works
The peroxisomal membrane protein Pex13p shows a novel mode of SH3 interaction
Barnett, P., Bottger, G., Klein, A. T. J., Tabak, H. F., & Distel, B. (2000) · The EMBO Journal · doi:10.1093/emboj/19.23.6382 · Cited by 89 · see citing works
Improved Scoring of Ligand−Protein Interactions Using OWFEG Free Energy Grids
Pearlman, D. A., & Charifson, P. S. (2001) · Journal of Medicinal Chemistry · doi:10.1021/jm000375v · Cited by 89 · see citing works
Similarity-driven flexible ligand docking
Fradera, X., Knegtel, R. M. A., & Mestres, J. (2000) · Proteins Structure Function and Bioinformatics · doi:10.1002/1097-0134(20000901)40:4 · Cited by 89 · see citing works
Protein structural alignments and functional genomics
Irving, J. A., Whisstock, J. C., & Lesk, A. M. (2001) · Proteins Structure Function and Bioinformatics · doi:10.1002/1097-0134(20010215)42:3 · Cited by 87 · see citing works
Bonneau, R., Strauss, C. E. M., & Baker, D. A. (2001) · Proteins Structure Function and Bioinformatics · doi:10.1002/1097-0134(20010401)43:1 · Cited by 82 · see citing works
Computer simulations of protein folding by targeted molecular dynamics
Ferrara, P., Apostolakis, J., & Caflisch, A. (2000) · Proteins Structure Function and Bioinformatics · doi:10.1002/(sici)1097-0134(20000515)39:3 · Cited by 78 · see citing works
Structural response to mutation at a protein-protein interface 1 1Edited by R. Huber
Vaughan, C. K., Buckle, A. M., & Fersht, A. R. (1999) · Journal of Molecular Biology · doi:10.1006/jmbi.1998.2559 · Cited by 72 · see citing works
Functional Significance of the β-Hairpin in the Insecticidal Neurotoxin ω-Atracotoxin-Hv1a
Tedford, H. W., Fletcher, J. I., & King, G. F. (2001) · Journal of Biological Chemistry · doi:10.1074/jbc.m102199200 · Cited by 72 · see citing works
Lysine 322 in the human IgG3 CH2 domain is crucial for antibody dependent complement activation
Thommesen, J. E., Michaelsen, T. E., Løset, G. Å., Sandlie, I., & Brekke, O. H. (2000) · Molecular Immunology · doi:10.1016/s0161-5890(01)00010-4 · Cited by 71 · see citing works
Single Amino Acid Substitutions in κ-Conotoxin PVIIA Disrupt Interaction with the Shaker K+ Channel
Jacobsen, R. B., Koch, E. D., Lange-Malecki, B., et al. (2000) · Journal of Biological Chemistry · doi:10.1074/jbc.c900990199 · Cited by 60 · see citing works
Identification of the ligand-binding site of the BMP type IA receptor for BMP-4
Hatta, T., Konishi, H., Katoh, E., Natsume, T., Ueno, N., Kobayashi, Y., & Yamazaki, T. (2000) · Biopolymers · doi:10.1002/1097-0282(2000)55:5 · Cited by 54 · see citing works
Shapiro, R., Ruiz‐Gutierrez, M., & Chen, C. (2000) · Journal of Molecular Biology · doi:10.1006/jmbi.2000.4075 · Cited by 51 · see citing works
Nakamura, G. R., Starovasnik, M. A., Reynolds, M. E., & Lowman, H. B. (2001) · Biochemistry · doi:10.1021/bi0109360 · Cited by 50 · see citing works
Epitope mapping for four monoclonal antibodies against human plasminogen activator inhibitor type‐1
Wind, T., Jensen, M. A., & Andreasen, P. A. (2001) · European Journal of Biochemistry · doi:10.1046/j.1432-1327.2001.2680041095.x · Cited by 49 · see citing works
Piehler, J., & Schreiber, G. (2001) · Analytical Biochemistry · doi:10.1006/abio.2000.4920 · Cited by 49 · see citing works
May the driving force be with you--whatever it is.
Cavanagh, J., & Akke, M. (2000) · Nature Structural Biology · doi:10.1038/71202 · Cited by 42 · see citing works
Allman, K., Page, K. M., CURTIS, C. A. M., et al. (2000) · Molecular Pharmacology · doi:10.1124/mol.58.1.175 · Cited by 41 · see citing works
Skelton, N. J., Chen, Y. M., Dubree, N., Quan, C. L., Jackson, D. Y., Cochran, A. G., Zobel, K., Deshayes, K., Baca, M., Pisabarro, M. T., et al. (2001) · Biochemistry · doi:10.1021/bi0103866 · Cited by 40 · see citing works
Clark, K. D., Volkman, B. F., Thoetkiattikul, H., et al. (2001) · Journal of Biological Chemistry · doi:10.1074/jbc.m105235200 · Cited by 27 · see citing works
Evaluation of protein–protein association energies by free energy perturbation calculations
Brandsdal, B. O., & Smalås, A. O. (2000) · Protein Engineering Design and Selection · doi:10.1093/protein/13.4.239 · Cited by 25 · see citing works
Theoretical Studies Suggest a New Antifolate as a More Potent Inhibitor of Thymidylate Synthase
Lee, T., & Kollman, P. A. (2000) · Journal of the American Chemical Society · doi:10.1021/ja9925554 · Cited by 19 · see citing works
Scott, W. N., McFerran, N. V., Harriott, P., Walker, B., & Nelson, J. (2000) · Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology · doi:10.1016/s0167-4838(00)00120-5 · Cited by 15 · see citing works
Structural consequences of an amino acid deletion in the B1 domain of protein G
O’Neil, K. T., Bach, A., & DeGrado, W. F. (2000) · Proteins Structure Function and Bioinformatics · doi:10.1002/1097-0134(20001115)41:3 · Cited by 11 · see citing works
Tikhomirov, O., & Thomas, J. W. (2000) · The Journal of Immunology · doi:10.4049/jimmunol.165.7.3876 · Cited by 10 · see citing works
Crystal structure of tropomyosin at 7 �ngstroms resolution
Whitby, F. G., & Phillips, G. N., Jr. (2000) · Proteins Structure Function and Bioinformatics · doi:10.1002/(sici)1097-0134(20000101)38:1
Flagged first, then by citations
Cited by
Sorted by citations
Small-molecule inhibitors of protein–protein interactions: progressing towards the dream
Arkin, M. R., & Wells, J. A. (2004) · Nature Reviews Drug Discovery · doi:10.1038/nrd1343 · Cited by 1,705 · see citing works
Hot spots—A review of the protein–protein interface determinant amino‐acid residues
Moreira, I. S., Fernandes, P. A., & Ramos, M. J. (2007) · Proteins: Structure, Function, and Bioinformatics · doi:10.1002/prot.21396 · Cited by 760 · see citing works
Ma, B., Elkayam, T., Wolfson, H., & Nussinov, R. (2003) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1030237100 · Cited by 595 · see citing works
Keskin, O., Ma, B., & Nussinov, R. (2005) · Journal of Molecular Biology · doi:10.1016/j.jmb.2004.10.077 · Cited by 519 · see citing works
Tethering: Fragment-Based Drug Discovery
Erlanson, D. A., Wells, J. A., & Braisted, A. C. (2004) · Annual Review of Biophysics and Biomolecular Structure · doi:10.1146/annurev.biophys.33.110502.140409 · Cited by 385 · see citing works
Binding MOAD (Mother Of All Databases)
Hu, L., Benson, M. L., Smith, R. D., Lerner, M. G., & Carlson, H. A. (2005) · Proteins: Structure, Function, and Bioinformatics · doi:10.1002/prot.20512 · Cited by 328 · see citing works
β-Hairpin Peptidomimetics: Design, Structures and Biological Activities
Robinson, J. A. (2008) · Accounts of Chemical Research · doi:10.1021/ar700259k · Cited by 321 · see citing works
Li, W., Asokan, A., Wu, Z., Van Dyke, T., DiPrimio, N., Johnson, J. S., Govindaswamy, L., Agbandje-McKenna, M., Leichtle, S., Eugene Redmond Jr, D., et al. (2008) · Molecular Therapy · doi:10.1038/mt.2008.100 · Cited by 261 · see citing works
Birtalan, S., Zhang, Y., Fellouse, F. A., Shao, L., Schaefer, G., & Sidhu, S. S. (2008) · Journal of Molecular Biology · doi:10.1016/j.jmb.2008.01.093 · Cited by 244 · see citing works
Mandal, S. M., Dey, S., Mandal, M., Sarkar, S., Maria-Neto, S., & Franco, O. L. (2009) · Peptides · doi:10.1016/j.peptides.2008.12.001 · Cited by 136 · see citing works
Protein–protein interactions as targets for small-molecule therapeutics in cancer
White, A. W., Westwell, A. D., & Brahemi, G. (2008) · Expert Reviews in Molecular Medicine · doi:10.1017/s1462399408000641 · Cited by 122 · see citing works
Lise, S., Archambeau, C., Pontil, M., & Jones, D. T. (2009) · BMC Bioinformatics · doi:10.1186/1471-2105-10-365 · Cited by 107 · see citing works
Targeting Protein–Protein Interactions and Fragment-Based Drug Discovery
Valkov, E., Sharpe, T., Marsh, M., Greive, S., & Hyvönen, M. (2011) · Topics in Current Chemistry · doi:10.1007/128_2011_265 · Cited by 107 · see citing works
A credit-card library approach for disrupting protein–protein interactions
Xu, Y., Shi, J., Yamamoto, N., Moss, J. A., Vogt, P. K., & Janda, K. D. (2006) · Bioorganic & Medicinal Chemistry · doi:10.1016/j.bmc.2005.11.052 · Cited by 93 · see citing works
Geng, C., Vangone, A., Folkers, G. E., Xue, L. C., & Bonvin, A. M. J. J. (2019) · Proteins: Structure, Function, and Bioinformatics · doi:10.1002/prot.25630 · Cited by 82 · see citing works
Prism: Protein-Protein Interaction Prediction by Structural Matching
Keskin, O., Nussinov, R., & Gursoy, A. (2008) · Methods in Molecular Biology · doi:10.1007/978-1-59745-398-1_30 · Cited by 80 · see citing works
Cierpicki, T., & Grembecka, J. (2015) · Immunological Reviews · doi:10.1111/imr.12244 · Cited by 53 · see citing works
Golden, M. S., Cote, S. M., Sayeg, M., Zerbe, B. S., Villar, E. A., Beglov, D., Sazinsky, S. L., Georgiadis, R. M., Vajda, S., Kozakov, D., et al. (2013) · Journal of the American Chemical Society · doi:10.1021/ja400914z · Cited by 47 · see citing works
Hybrid Organic–Inorganic Inhibitors of a PDZ Interaction that Regulates the Endocytic Fate of CFTR
Kundu, R., Cushing, P. R., Popp, B. V., Zhao, Y., Madden, D. R., & Ball, Z. T. (2012) · Angewandte Chemie International Edition · doi:10.1002/anie.201202291 · Cited by 33 · see citing works
Bohnuud, T., Kozakov, D., & Vajda, S. (2014) · PLoS Computational Biology · doi:10.1371/journal.pcbi.1003872 · Cited by 27 · see citing works
Protein-Protein Interactions as Drug Targets
Wendt, M. D. (2012) · Topics in Medicinal Chemistry · doi:10.1007/978-3-642-28965-1_1 · Cited by 9 · see citing works
Xie, X., Sun, L., Pessetto, Z. Y., Zhao, Y., Zang, Z., Zhong, L., Wu, M., Su, Q., Gao, X., Zan, W., et al. (2013) · PLoS ONE · doi:10.1371/journal.pone.0078042 · Cited by 8 · see citing works
- Authors
- DeLano, W. L.
- Affiliations
- Sunesis (United States) (US)
- Corresponding author
- DeLano, Warren L
- Venue
- Current Opinion in Structural Biology
- Publisher
- Elsevier BV
- Published
- 2002-02
- Volume
- 12
- Issue
- 1
- Pages
- 14-20
- ISSN
- 0959-440X (ISSN-L)
- OpenAlex
- W2026370952
- Cited by
- 767
- References
- 46
- Citations by year
- 2026: 14 · 2025: 10 · 2024: 23 · 2023: 19 · 2022: 30 · 2021: 11 · 2020: 24 · 2019: 34 · 2018: 31 · 2017: 18 · 2016: 25 · 2015: 27 · 2014: 43 · 2013: 43 · 2012: 62
- FWCI
- 2.27
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Current Opinion in Structural Biology (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
- Physical Sciences › Computer Science › Computational Theory and Mathematics › Computational Drug Discovery Methods
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Interaction Studies and Fluorescence Analysis
- Keywords
- protein-protein interfaces, alanine scanning, hot spot prediction, structural plasticity, molecular recognition, drug discovery
- MeSH terms
- Alanine, Binding Sites, Drug Therapy, Humans, Models, Molecular, Protein Binding, Thermodynamics
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review