TY  - JOUR
AU  - Dall, Elfriede
AU  - Stanojlovic, Vesna
AU  - Demir, Fatih
AU  - Briza, Peter
AU  - Dahms, Sven O.
AU  - Huesgen, Pitter F.
AU  - Cabrele, Chiara
AU  - Brandstetter, Hans
TI  - The Peptide Ligase Activity of Human Legumain Depends on Fold Stabilization and Balanced Substrate Affinities
JO  - ACS catalysis
VL  - 11
IS  - 19
SN  - 2155-5435
CY  - Washington, DC
PB  - ACS
M1  - FZJ-2021-03503
SP  - 11885 - 11896
PY  - 2021
AB  - Protein modification by enzymatic breaking and forming of peptide bonds significantly expands the repertoire of genetically encoded protein sequences. The dual protease-ligase legumain exerts the two opposing activities within a single protein scaffold. Primarily localized to the endolysosomal system, legumain represents a key enzyme in the generation of antigenic peptides for subsequent presentation on the MHCII complex. Here we show that human legumain catalyzes the ligation and cyclization of linear peptides at near-neutral pH conditions, where legumain is intrinsically unstable. Conformational stabilization significantly enhanced legumain’s ligase activity, which further benefited from engineering the prime substrate recognition sites for improved affinity. Additionally, we provide evidence that specific legumain activation states allow for differential regulation of its activities. Together these results set the basis for engineering legumain proteases and ligases with applications in biotechnology and drug development.
LB  - PUB:(DE-HGF)16
C6  - 34621593
UR  - <Go to ISI:>//WOS:000704700800008
DO  - DOI:10.1021/acscatal.1c02057
UR  - https://juser.fz-juelich.de/record/894963
ER  -