Inâ
Situ Cyclization of Native Proteins: Structure-Based Design of a Bicyclic Enzyme.
Angew Chem Int Ed Engl
; 57(35): 11164-11170, 2018 08 27.
Article
in En
| MEDLINE
| ID: mdl-29847004
ABSTRACT
Increased tolerance of enzymes towards thermal and chemical stress is required for many applications and can be achieved by macrocyclization of the enzyme resulting in the stabilizing of its tertiary structure. Thus far, macrocyclization approaches utilize a very limited structural diversity, which complicates the design process. Herein, we report an approach that enables cyclization through the installation of modular crosslinks into native proteins composed entirely of proteinogenic amino acids. Our stabilization procedure involves the introduction of three surface-exposed cysteine residues, which are reacted with a triselectrophile, resulting in the inâ
situ cyclization of the protein (INCYPRO). A bicyclic version of sortaseâ
A was designed that exhibits increased tolerance towards thermal as well as chemical denaturation, and proved to be efficient in protein labeling under denaturing conditions. In addition, we applied INCYPRO to the KIX domain, resulting in up to 24 °C increased thermal stability.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Staphylococcus aureus
/
Bacterial Proteins
/
Cysteine Endopeptidases
/
Aminoacyltransferases
/
Cross-Linking Reagents
/
Cysteine
Limits:
Animals
/
Humans
Language:
En
Journal:
Angew Chem Int Ed Engl
Year:
2018
Type:
Article
Affiliation country:
Netherlands