title: Porcine amino peptidase N domain VII has critical role in binding and entry of porcine epidemic diarrhea virus.
abstract: Porcine epidemic diarrhea virus (PEDV) infects swine intestinal cells causing enteric disease. Research has shown that the entry into these cells is through porcine aminopeptidase N (pAPN) receptor. To gain insights into mechanisms of PEDV-pAPN interactions, the present study aimed at identifying the domain that is critical for PEDV binding. To this end, NIH3T3 cell lines constitutively expressing pAPN or pAPN mutants were generated. The mutants were; domain VII deletion mutant and domains IV-VI deletion mutant. In the latter, domain VII was linked to the transmembrane segment through domain III. Results showed PEDV infection was restricted to pAPN and pAPN domain VII expressing NIH3T3 cells. Further, reducing PEDV titre 10 fold resulted in 37.8% decrease in foci indicating positive correlation. A time course test at 12, 24, 36, 48 and 60h showed that foci increased 6 fold in the overall time range. Also, PEDV harvested from pAPN or domain VII expressing NIH3T3 cells was induced indirect plaques in Vero cells confirming successful entry and replication. Collectively, our results demonstrate that PEDV recognizes pAPN and that the main interactive point is lodged within domain VII of the pAPN. These findings are important for therapeutic development as well as creating a platform for future studies on PEDV.
authors.affiliation: Laboratory of Infectious Diseases, College of Veterinary Medicine, Chungnam National University, 220 Gungdong, Yuseong, Daejeon, 305-764, Republic of Korea.
authors.forename: Anthony Ndirangu
authors.initials: AN
authors.lastname: Kamau
authors.affiliation: Laboratory of Infectious Diseases, College of Veterinary Medicine, Chungnam National University, 220 Gungdong, Yuseong, Daejeon, 305-764, Republic of Korea.
authors.forename: Jung-Eun
authors.initials: JE
authors.lastname: Park
authors.affiliation: Department of Microbiology & Molecular Biology College of Bioscience & Biotechnology, 220 Gungdong, Yuseong, Daejeon, 305-764, Republic of Korea.
authors.forename: Eui-Soon
authors.initials: ES
authors.lastname: Park
authors.affiliation: Department of Microbiology & Molecular Biology College of Bioscience & Biotechnology, 220 Gungdong, Yuseong, Daejeon, 305-764, Republic of Korea.
authors.forename: Jung-Eun
authors.initials: JE
authors.lastname: Yu
authors.affiliation: Department of Microbiology & Molecular Biology College of Bioscience & Biotechnology, 220 Gungdong, Yuseong, Daejeon, 305-764, Republic of Korea.
authors.forename: Jaerang
authors.initials: J
authors.lastname: Rho
authors.affiliation: Laboratory of Infectious Diseases, College of Veterinary Medicine, Chungnam National University, 220 Gungdong, Yuseong, Daejeon, 305-764, Republic of Korea; Research Institute of Veterinary Medicine, 220 Gungdong, Yuseong, Daejeon, 305-764, Republic of Korea. Electronic address: shin0089@cnu.ac.kr.
authors.forename: Hyun-Jin
authors.initials: HJ
authors.lastname: Shin
mesh_terms: D000818:Animals; D018826:CD13 Antigens / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D018352:Coronavirus Infections / Q000662:veterinary*; D015870:Gene Expression; D057809:HEK293 Cells; D006801:Humans; D051379:Mice; D041681:NIH 3T3 Cells; D053485:Porcine epidemic diarrhea virus / Q000502:physiology*; D000072417:Protein Domains; D011991:Receptors, Virus / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013552:Swine; D013553:Swine Diseases / Q000378:metabolism* / Q000821:virology*; D053585:Virus Attachment; D053586:Virus Internalization; D014779:Virus Replication
chemical_list: D011991:Receptors, Virus; D018826:CD13 Antigens
keywords: Binding and entry; Domain VII; Pathogenesis; Porcine APN; Porcine epidemic diarrhea virus