A Novel Purification Procedure for Active Recombinant Human DPP4 and the Inability of DPP4 to Bind SARS-CoV-2

Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry
Cecy XiMark D Gorrell

Abstract

Proteases catalyse irreversible posttranslational modifications that often alter a biological function of the substrate. The protease dipeptidyl peptidase 4 (DPP4) is a pharmacological target in type 2 diabetes therapy primarily because it inactivates glucagon-like protein-1. DPP4 also has roles in steatosis, insulin resistance, cancers and inflammatory and fibrotic diseases. In addition, DPP4 binds to the spike protein of the MERS virus, causing it to be the human cell surface receptor for that virus. DPP4 has been identified as a potential binding target of SARS-CoV-2 spike protein, so this question requires experimental investigation. Understanding protein structure and function requires reliable protocols for production and purification. We developed such strategies for baculovirus generated soluble recombinant human DPP4 (residues 29-766) produced in insect cells. Purification used differential ammonium sulphate precipitation, hydrophobic interaction chromatography, dye affinity chromatography in series with immobilised metal affinity chromatography, and ion-exchange chromatography. The binding affinities of DPP4 to the SARS-CoV-2 full-length spike protein and its receptor-binding domain (RBD) were measured using surface p...Continue Reading

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Citations

Feb 3, 2021·Archivum Immunologiae Et Therapiae Experimentalis·Alicja Krejner-BieniasTomasz Grzela
Mar 17, 2021·Biochemical Pharmacology·R YazbeckC A Abbott
Oct 29, 2021·Molecular Systems Biology·Xiaonan LiuMarkku Varjosalo

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Datasets Mentioned

BETA
M80536
AHX00711.1

Methods Mentioned

BETA
glycosylation
size-exclusion chromatography
ELISA
surface plasmon resonance
ion-exchange chromatography
chip
tandem dye affinity chromatography
restriction digest
transfection
affinity purification

Software Mentioned

Graphpad Prism
BIAevaluation
PyMOL

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