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Identification of residues on human receptor DPP4 critical for MERS-CoV binding and entry
Wenfei Song1, Ying Wang2, Nianshuang Wang1
1Ministry of Education Key Laboratory of Protein Science, Center for Structural Biology, School of Life Sciences, Tsinghua University, Beijing 100084, China.
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.
Insights
Researchers identified key residues on the human receptor dipeptidyl peptidase 4 (hDPP4) crucial for Middle East respiratory syndrome coronavirus (MERS-CoV) spike protein binding and viral entry, offering insights into MERS-CoV tropism.
Area of Science:
- Virology
- Structural Biology
- Biochemistry
Background:
- Middle East respiratory syndrome coronavirus (MERS-CoV) utilizes its spike glycoprotein's receptor binding domain (RBD) to bind human dipeptidyl peptidase 4 (hDPP4), initiating host cell infection.
- Understanding the molecular interactions at the RBD-hDPP4 interface is critical for comprehending MERS-CoV entry mechanisms.
Purpose of the Study:
- To identify specific amino acid residues on hDPP4 that are essential for MERS-CoV RBD binding and subsequent viral entry.
- To elucidate the structural and functional significance of these critical residues at the molecular level.
Main Methods:
- Analysis of a panel of engineered hDPP4 mutants with altered residues at the RBD-hDPP4 interface.
- Surface plasmon resonance (SPR) binding assays to quantify the binding affinity between MERS-CoV RBD and hDPP4 mutants.
- Pseudovirus infection assays to assess the impact of hDPP4 mutations on viral entry efficiency.
Main Results:
- Several mutated residues located at the RBD-hDPP4 interface were found to be critical for both RBD binding and viral entry.
- Alterations in these residues potentially affected the polarity, hydrophobic, or hydrophilic properties of hDPP4, disrupting the interaction with MERS-CoV RBD.
- SPR and pseudovirus assays confirmed the importance of these interface residues in mediating MERS-CoV infection.
Conclusions:
- Atomic-level insights into the interaction between MERS-CoV RBD and hDPP4 have been provided.
- The identified critical residues offer a potential explanation for the cellular and species tropism observed in MERS-CoV infections.
- This study lays the groundwork for developing targeted interventions against MERS-CoV entry.
