Hot regions in protein-protein interactions: The organization and contribution of structurally conserved hot spot residues

Ozlem Keskin, Buyong Ma, Ruth Nussinov

Research output: Contribution to journalArticlepeer-review

441 Scopus citations


Structurally conserved residues at protein-protein interfaces correlate with the experimental alanine-scanning hot spots. Here, we investigate the organization of these conserved, computational hot spots and their contribution to the stability of protein associations. We find that computational hot spots are not homogeneously distributed along the protein interfaces; rather they are clustered within locally tightly packed regions. Within the dense clusters, they form a network of interactions and consequently their contributions to the stability of the complex are cooperative; however the contributions of independent clusters are additive. This suggests that the binding free energy is not a simple summation of the single hot spot residue contributions. As expected, around the hot spot residues we observe moderately conserved residues, further highlighting the crucial role of the conserved interactions in the local densely packed environment. The conserved occurrence of these organizations suggests that they are advantageous for protein-protein associations. Interestingly, the total number of hydrogen bonds and salt bridges contributed by hot spots is as expected. Thus, H-bond forming residues may use a "hot spot for water exclusion" mechanism. Since conserved residues are located within highly packed regions, water molecules are easily removed upon binding, strengthening electrostatic contributions of charge-charge interactions. Hence, the picture that emerges is that protein-protein associations are optimized locally, with the clustered, networked, highly packed structurally conserved residues contributing dominantly and cooperatively to the stability of the complex. When addressing the crucial question of "what are the preferred ways of proteins to associate", these findings point toward a critical involvement of hot regions in protein-protein interactions.

Original languageEnglish
Pages (from-to)1281-1294
Number of pages14
JournalJournal of Molecular Biology
Issue number5
StatePublished - 4 Feb 2005


FundersFunder number
National Institutes of HealthNO1-CO-12400
National Cancer InstituteZ01BC010441
Israel Academy of Sciences and Humanities
Israel Science Foundation


    • hot spots
    • protein-protein interactions
    • residue conservation
    • residue cooparativity
    • residue networks


    Dive into the research topics of 'Hot regions in protein-protein interactions: The organization and contribution of structurally conserved hot spot residues'. Together they form a unique fingerprint.

    Cite this