Abstract
Acyl aminoacyl peptidases are two-domain proteins composed by a C-terminal catalytic α/β-hydrolase domain and by an N-terminal β-propeller domain connected through a structural element that is at the N-terminus in sequence but participates in the 3D structure of the C-domain. We investigated about the structural and functional interplay between the two domains and the bridge structure (in this case a single helix named α1-helix) in the cold-adapted enzyme from Sporosarcina psychrophila (SpAAP) using both protein variants in which entire domains were deleted and proteins carrying substitutions in the α1-helix. We found that in this enzyme the inter-domain connection dramatically affects the stability of both the whole enzyme and the β-propeller. The α1-helix is required for the stability of the intact protein, as in other enzymes of the same family; however in this psychrophilic enzyme only, it destabilizes the isolated β-propeller. A single charged residue (E10) in the α1-helix plays a major role for the stability of the whole structure. Overall, a strict interaction of the SpAAP domains seems to be mandatory for the preservation of their reciprocal structural integrity and may witness their co-evolution.
| Original language | English |
|---|---|
| Article number | e56254 |
| Journal | PloS one |
| Volume | 8 |
| Issue number | 2 |
| Number of pages | 11 |
| ISSN | 1932-6203 |
| DOIs | |
| Publication status | Published - 15 Feb 2013 |
| Externally published | Yes |
Keywords
- Adaptation, Physiological
- Catalytic Domain
- Cloning, Molecular
- Cold Temperature
- Enzyme Stability
- Molecular Dynamics Simulation
- Mutagenesis
- Peptide Hydrolases
- Protein Structure, Secondary
- Protein Structure, Tertiary
- Recombinant Proteins
- Sequence Deletion
- Solubility
- Sporosarcina