Fast and faster: a designed variant of the B-domain of protein A folds in 3 microsec.
We have introduced the mutation glycine 29 to alanine, designed to increase the rate of protein folding, into the B-domain of protein A (BdpA). From NMR lineshape analysis, we find the G29A mutation increases the folding rate constant by threefold; the folding time is 3 microsec. Although wild-type BdpA folds extremely fast, simple-point mutations can still speed up the folding; thus, the folding rate is not evolutionarily maximized. The short folding time of G29A BdpA (the shortest time yet reported) makes it an attractive candidate for an all-atom molecular dynamics simulation that could potentially show a complete folding reaction starting from an extended chain. We also constructed a fluorescent variant of BdpA by mutating phenylalanine 13 to tryptophan, allowing fluorescence-based time-resolved temperature-jump measurements. Temperature jumps and NMR complement each other, and give a very complete picture of the folding kinetics.
Duke Scholars
Altmetric Attention Stats
Dimensions Citation Stats
Published In
DOI
ISSN
Publication Date
Volume
Issue
Start / End Page
Location
Related Subject Headings
- Staphylococcal Protein A
- Protein Structure, Tertiary
- Protein Folding
- Protein Denaturation
- Models, Molecular
- Magnetic Resonance Spectroscopy
- Kinetics
- Biophysics
- Amino Acid Substitution
- 3404 Medicinal and biomolecular chemistry
Citation
Published In
DOI
ISSN
Publication Date
Volume
Issue
Start / End Page
Location
Related Subject Headings
- Staphylococcal Protein A
- Protein Structure, Tertiary
- Protein Folding
- Protein Denaturation
- Models, Molecular
- Magnetic Resonance Spectroscopy
- Kinetics
- Biophysics
- Amino Acid Substitution
- 3404 Medicinal and biomolecular chemistry