5Z65

Crystal structure of porcine aminopeptidase N ectodomain in functional form

Summary for 5Z65

• Entry DOI: 10.2210/pdb5z65/pdb
• Related PRD ID: PRD_900017
• Descriptor: Aminopeptidase, 2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose, 2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose, ... (6 entities in total)
• Functional Keywords: hydrolase
• Biological source: Sus scrofa (Pig)
• Total number of polymer chains: 1
• Total formula weight: 108207.08

Authors

Sun, Y.G.,Li, R.,Jiang, L.G.,Qiao, S.L.,Zhang, G.P. (deposition date: 2018-01-22, release date: 2019-01-02, Last modification date: 2023-11-22)

Primary citation

Sun, Y.G.,Li, R.,Jiang, L.,Qiao, S.,Zhi, Y.,Chen, X.X.,Xie, S.,Wu, J.,Li, X.,Deng, R.,Zhang, G.
Characterization of the interaction between recombinant porcine aminopeptidase N and spike glycoprotein of porcine epidemic diarrhea virus.
Int. J. Biol. Macromol., 117:704-712, 2018

PubMed Abstract: Porcine epidemic diarrhea (PED) has caused huge economic losses to the global pork industry. Infection by its causative agent PED virus (PEDV), an Alpha-coronavirus, was previously proven to be mediated by its spike (S) glycoprotein and a cellular receptor porcine aminopeptidase N (pAPN). Interestingly, some recent studies have indicated that pAPN is not a functional receptor for PEDV. To date, there is a lack of a direct evidence for the interaction between pAPN and PEDV S protein in vitro. Here, we prepared pAPN ectodomain and the truncated variants of PEDV S protein in Drosophila S2 cells. These recombinant proteins were homogeneous after purification by metal-affinity and size-exclusion chromatography. We then assayed the purified target proteins through immunogenicity tests, PEDV binding interference assays, circular dichroism (CD) measurements, pAPN activity assay and structural determination, demonstrating that they were biologically functional. Finally, we characterized their interactions by gel filtration chromatography, native-polyacrylamide gel electrophoresis (PAGE) and surface plasmon resonance (SPR) analyses. The results showed that their affinities were too low to form complexes, which suggest that pAPN may be controversial as the genuine receptor for PEDV. Therefore, further research needs to be carried out to elucidate the interaction between PEDV and its genuine receptor.

PubMed: 29802920
DOI: 10.1016/j.ijbiomac.2018.05.167

Experimental method

X-RAY DIFFRACTION (2.65 Å)