Journal
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Volume 102, Issue 36, Pages 12724-12729Publisher
NATL ACAD SCIENCES
DOI: 10.1073/pnas.0506124102
Keywords
adaptor protein; protein engineering; SspB
Categories
Funding
- NCRR NIH HHS [P41 RR015301, RR-15301] Funding Source: Medline
- NIAID NIH HHS [R01 AI016892, AI-15706, R37 AI015706, AI-16892, R01 AI015706] Funding Source: Medline
Ask authors/readers for more resources
Protein-protein interactions can be designed computationally by using positive strategies that maximize the stability of the desired structure and/or by negative strategies that seek to destabilize competing states. Here, we compare the efficacy of these methods in reengineering a protein homodimer into a heterodimer. The stability-design protein (positive design only) was experimentally more stable than the specificity-design heterodimer (positive and negative design). By contrast, only the specificity-design protein assembled as a homogenous heterodimer in solution, whereas the stability-design protein formed a mixture of homodimer and heteroclimer species. The experimental stabilities of the engineered proteins correlated roughly with their calculated Stabilities, and the crystal structure of the specificity-design heterodimer showed most of the predicted side-chain packing interactions and a mainchain conformation indistinguishable from the wild-type structure. These results indicate that the design simulations capture important features of both stability and structure and demonstrate that negative design can be critical for attaining specificity when competing states are close in structure space.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available