Structural insights into thrombolytic activity of destabilase from medicinal leech.
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
24 04 2023
24 04 2023
Historique:
received:
14
01
2023
accepted:
28
03
2023
medline:
26
4
2023
pubmed:
25
4
2023
entrez:
24
04
2023
Statut:
epublish
Résumé
Destabilase from the medical leech Hirudo medicinalis belongs to the family of i-type lysozymes. It has two different enzymatic activities: microbial cell walls destruction (muramidase activity), and dissolution of the stabilized fibrin (isopeptidase activity). Both activities are known to be inhibited by sodium chloride at near physiological concentrations, but the structural basis remains unknown. Here we present two crystal structures of destabilase, including a 1.1 Å-resolution structure in complex with sodium ion. Our structures reveal the location of sodium ion between Glu34/Asp46 residues, which were previously recognized as a glycosidase active site. While sodium coordination with these amino acids may explain inhibition of the muramidase activity, its influence on previously suggested Ser49/Lys58 isopeptidase activity dyad is unclear. We revise the Ser49/Lys58 hypothesis and compare sequences of i-type lysozymes with confirmed destabilase activity. We suggest that the general base for the isopeptidase activity is His112 rather than Lys58. pKa calculations of these amino acids, assessed through the 1 μs molecular dynamics simulation, confirm the hypothesis. Our findings highlight the ambiguity of destabilase catalytic residues identification and build foundations for further research of structure-activity relationship of isopeptidase activity as well as structure-based protein design for potential anticoagulant drug development.
Identifiants
pubmed: 37095116
doi: 10.1038/s41598-023-32459-x
pii: 10.1038/s41598-023-32459-x
pmc: PMC10126035
doi:
Substances chimiques
fibrin destabilase
EC 3.4.99.-
Muramidase
EC 3.2.1.17
Endopeptidases
EC 3.4.-
Fibrinolytic Agents
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
6641Informations de copyright
© 2023. The Author(s).
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