title: Identification of residues on human receptor DPP4 critical for MERS-CoV binding and entry.
abstract: Middle East respiratory syndrome coronavirus (MERS-CoV) infects host cells through binding the receptor binding domain (RBD) on its spike glycoprotein to human receptor dipeptidyl peptidase 4 (hDPP4). Here, we report identification of critical residues on hDPP4 for RBD binding and virus entry through analysis of a panel of hDPP4 mutants. Based on the RBD-hDPP4 crystal structure we reported, the mutated residues were located at the interface between RBD and hDPP4, which potentially changed the polarity, hydrophobic or hydrophilic properties of hDPP4, thereby interfering or disrupting their interaction with RBD. Using surface plasmon resonance (SPR) binding analysis and pseudovirus infection assay, we showed that several residues in hDPP4-RBD binding interface were important on hDPP4-RBD binding and viral entry. These results provide atomic insights into the features of interactions between hDPP4 and MERS-CoV RBD, and also provide potential explanation for cellular and species tropism of MERS-CoV infection.
authors.affiliation: Ministry of Education Key Laboratory of Protein Science, Center for Structural Biology, School of Life Sciences, Tsinghua University, Beijing 100084, China.
authors.forename: Wenfei
authors.initials: W
authors.lastname: Song
authors.affiliation: Comprehensive AIDS Research Center, Research Center for Public Health, School of Medicine, Tsinghua University, Beijing 100084, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Ministry of Education Key Laboratory of Protein Science, Center for Structural Biology, School of Life Sciences, Tsinghua University, Beijing 100084, China.
authors.forename: Nianshuang
authors.initials: N
authors.lastname: Wang
authors.affiliation: Ministry of Education Key Laboratory of Protein Science, Center for Structural Biology, School of Life Sciences, Tsinghua University, Beijing 100084, China.
authors.forename: Dongli
authors.initials: D
authors.lastname: Wang
authors.affiliation: Comprehensive AIDS Research Center, Research Center for Public Health, School of Medicine, Tsinghua University, Beijing 100084, China.
authors.forename: Jianying
authors.initials: J
authors.lastname: Guo
authors.affiliation: Comprehensive AIDS Research Center, Research Center for Public Health, School of Medicine, Tsinghua University, Beijing 100084, China.
authors.forename: Lili
authors.initials: L
authors.lastname: Fu
authors.affiliation: Comprehensive AIDS Research Center, Research Center for Public Health, School of Medicine, Tsinghua University, Beijing 100084, China. Electronic address: shixuanlingsk@tsinghua.edu.cn.
authors.forename: Xuanling
authors.initials: X
authors.lastname: Shi
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D018819:Dipeptidyl Peptidase 4 / Q000235:genetics / Q000378:metabolism*; D015967:Gene Expression Regulation, Viral / Q000502:physiology*; D006801:Humans; D007313:Insecta; D065207:Middle East Respiratory Syndrome Coronavirus / Q000502:physiology*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D053586:Virus Internalization
chemical_list: C042807:DPP4 protein, human; D018819:Dipeptidyl Peptidase 4
keywords: Amino-acid residue substitution; MERS-CoV; RBD; hDPP4