Generation and analysis of proline mutants in protein G.


The pyrrolidine ring of the amino acid proline reduces the conformational freedom of the protein backbone in its unfolded form and thus enhances protein stability. The strategy of inserting proline into regions of the protein where it does not perturb the structure has been utilized to stabilize many different proteins including enzymes. However, most of these efforts have been based on trial and error, rather than rational design. Here, we try to understand proline's effect on protein stability by introducing proline mutations into various regions of the B1 domain of Streptococcal protein G. We also applied the Optimization of Rotamers By Iterative Techniques computational protein design program, using two different solvation models, to determine the extent to which it could predict the stabilizing and destabilizing effects of prolines. Use of a surface area dependent solvation model resulted in a modest correlation between the experimental free energy of folding and computed energies; on the other hand, use of a Gaussian solvent exclusion model led to significant positive correlation. Including a backbone conformational entropy term to the computational energies increases the statistical significance of the correlation between the experimental stabilities and both solvation models. Study holds ProTherm entries: 21957, 21958, 21959, 21960, 21961, 21962, 21963, 21964, 21965, 21966, 21967, 21968, 21969, 21970, 21971, 21972, 21973, 21974, 21975, 21976, 21977, 21978, 21979, 21980 Extra Details: B1 domain proline; protein; design; protein G; protein stability

Submission Details

ID: 9KEVemSb

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:53 p.m.

Version: 1

Publication Details
Choi EJ;Mayo SL,Protein Eng. Des. Sel. (2006) Generation and analysis of proline mutants in protein G. PMID:16549401
Additional Information

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Immunoglobulin G-binding protein G P06654 SPG1_STRSG
100.0 Immunoglobulin G-binding protein G P19909 SPG2_STRSG