The Feline calicivirus Leader of the Capsid (LC) Protein Contains a Putative Transmembrane Domain, Binds to the Cytoplasmic Membrane, and Exogenously Permeates Cells
Abstract
:1. Introduction
2. Materials and Methods
2.1. Prediction and Validation of the Tertiary Structure of the WT and C40A Mutant LC Proteins
2.2. Prediction of the Transmembrane Domain of the WT and C40A Mutant LC Proteins
2.3. Cell Culture
2.4. Plasmid Purification and Transfection
2.5. Recombinant Protein Expression and Purification
2.6. In Vitro Exogenous Interaction of the LC Protein with the Plasmatic Membrane of CrFK Cells
2.7. Cell Permeation Assays
2.8. SDS-PAGE Western Blot Analysis
3. Results
3.1. Validation of the Predicted Tertiary Structure of the Wild Type and the C40A Mutant LC Proteins from the FCV
3.2. Putative Wild-Type and C40A Mutant LC Protein Tertiary Structures Possess Intramolecular Disulfide Bonds in Accordance with Experimental Data
3.3. The FCV LC Protein Predicted Structure Lacks a Defensin γ-Core Signature in Its CR-I But Has a Putative C-Terminal Transmembrane Domain
3.4. Modeling of FCV Wild Type and C40A Mutant LC Protein Oligomeric Forms by AlphaFold 2
3.5. The Purified Histag-LC Protein from FCV Associates with the CrFK Cells’ Cytoplasmic Membrane through Exogenous Interaction
3.6. The Purified Histag-LC Protein from FCV Permeates CrFK Cells’ Cytoplasmic Membrane through Exogenous Interaction and in a Disulfide Bond-Independent Formation
3.7. The Purified Histag-LC Protein from FCV Permeates the Cytoplasmic Membrane of the Human SKOV3 and the Murine ID8 Ovarian Cancer Cells through an Exogenous Interaction
3.8. The FCV LC Protein Expression in a Virus-Free System Induces Apoptosis in SKOV3 Cells
4. Discussion
Supplementary Materials
Author Contributions
Funding
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
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Peñaflor-Téllez, Y.; Escobar-Almazan, J.A.; Pérez-Ibáñez, C.; Miguel-Rodríguez, C.E.; Gómez de la Madrid, J.; Monge-Celestino, E.I.; Talamás-Rohana, P.; Gutiérrez-Escolano, A.L. The Feline calicivirus Leader of the Capsid (LC) Protein Contains a Putative Transmembrane Domain, Binds to the Cytoplasmic Membrane, and Exogenously Permeates Cells. Viruses 2024, 16, 1319. https://doi.org/10.3390/v16081319
Peñaflor-Téllez Y, Escobar-Almazan JA, Pérez-Ibáñez C, Miguel-Rodríguez CE, Gómez de la Madrid J, Monge-Celestino EI, Talamás-Rohana P, Gutiérrez-Escolano AL. The Feline calicivirus Leader of the Capsid (LC) Protein Contains a Putative Transmembrane Domain, Binds to the Cytoplasmic Membrane, and Exogenously Permeates Cells. Viruses. 2024; 16(8):1319. https://doi.org/10.3390/v16081319
Chicago/Turabian StylePeñaflor-Téllez, Yoatzin, Jesús Alejandro Escobar-Almazan, Carolina Pérez-Ibáñez, Carlos Emilio Miguel-Rodríguez, Jaury Gómez de la Madrid, Erick I. Monge-Celestino, Patricia Talamás-Rohana, and Ana Lorena Gutiérrez-Escolano. 2024. "The Feline calicivirus Leader of the Capsid (LC) Protein Contains a Putative Transmembrane Domain, Binds to the Cytoplasmic Membrane, and Exogenously Permeates Cells" Viruses 16, no. 8: 1319. https://doi.org/10.3390/v16081319