Biochemical and functional characterization of a new recombinant phospholipase A2 inhibitor from Crotalus durissus collilineatus snake serum release_bfds6uatrbekxh54bifan3pggq

by Sarah Natalie Cirilo Gimenes, Lorenz Aglas, Sabrina Wildner, Sara Huber, Ana Carolina Portella Silveira, Daiana Silva Lopes, Renata Santos Rodrigues, Luiz Ricardo Goulart, Peter Briza, Fatima Ferreira, Veridiana de Melo Rodrigues Ávila, Gabriele Gadermaier

Published in International Journal of Biological Macromolecules by Elsevier BV.

2020  

Abstract

Phospholipase A2 plays an important role in many diseases. Thus, the production of bioactive molecules, which can modulate PLA2 activity, became an important target for the pharmaceutical industry. Previously, we demonstrated the inhibitory and anti-angiogenic effect of γCdcPLI, the natural PLA2inhibitor from Crotalus durissus collilineatus. The aim of the present study was to recombinantly express the γCdcPLI inhibitor and analyze its biochemical and functional characteristics. Based on the amino acid sequence from the natural protein, we designed a synthetic gene for production of a non-tagged recombinant recγCdcPLI using the pHis-Parallel2 vector. To enable disulfide bond formation, protein expression was performed using E. coli Rosetta-gamiB. The protein was purified by anion and affinity chromatography with a yield of 5 mg/l. RecγCdcPLI showed similar secondary structure in CD and FTIR, revealing predominately β-strands. Analogous to the natural protein, recγCdcPLI was able to form oligomers of ~5.5 nm. The inhibitor was efficiently binding to PLA2 from honeybee (Kd = 1.48 μM) and was able to inhibit the PLA2 activity. Furthermore, it decreased the vessel formation in HUVEC cells, suggesting an anti-angiogenic potential. Heterologous production of recγCdcPLI is highly efficient and thus enables enhanced drug design for treatment of diseases triggered by PLA2 activity.
In text/plain format

Archived Content

There are no accessible files associated with this release. You could check other releases for this work for an accessible version.

"Dark" Preservation Only
Save Paper Now!

Know of a fulltext copy of on the public web? Submit a URL and we will archive it

Type  article-journal
Stage   published
Date   2020-07-28
Language   en ?
Journal Metadata
Not in DOAJ
In Keepers Registry
ISSN-L:  0141-8130
Work Entity
access all versions, variants, and formats of this works (eg, pre-prints)
Catalog Record
Revision: b431e9e2-89aa-4353-a92a-a1e5571b70d2
API URL: JSON