Specific Binding and Endocytosis of Liposomes to HEK293T Cells via Myrisoylated Pre-S1 Peptide Bound to Sodium Taurocholate Cotransporting Polypeptide
(1) Background: Sodium taurocholate cotransporting polypeptide (NTCP) functions as a key receptor for the hepatitis B virus (HBV) infection. Analyzing HBV and NTCP interaction is an important issue not only for basic research but also for the development of anti-HBV therapeutics. We developed here a...
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| Format: | Article |
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MDPI AG
2022-11-01
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| Series: | Vaccines |
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| Online Access: | https://www.mdpi.com/2076-393X/10/12/2050 |
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| author | Shuji Hinuma Kazuyo Fujita Shun’ichi Kuroda |
| author_facet | Shuji Hinuma Kazuyo Fujita Shun’ichi Kuroda |
| author_sort | Shuji Hinuma |
| collection | DOAJ |
| description | (1) Background: Sodium taurocholate cotransporting polypeptide (NTCP) functions as a key receptor for the hepatitis B virus (HBV) infection. Analyzing HBV and NTCP interaction is an important issue not only for basic research but also for the development of anti-HBV therapeutics. We developed here a novel model system to analyze the interaction of NTCP with liposomes instead of HBV. (2) Methods: Liposomal binding and endocytosis through NTCP in HEK293T cells were achieved by serial treatments of HEL293T cells transiently expressing NTCP-green fluorescence protein (GFP) fusion protein with a synthetic biotinylated pre-S1 peptide (Myr47-Bio) and streptavidin (SA) complex (i.e., Myr47-Bio+SA) followed by biotinylated liposomes. By this procedure, binding of [biotinylated liposomes]-[Myr47-Bio+SA]-[NTCP-GFP] was formed. (3) Results: Using this model system, we found that liposomal binding to NTCP on the cell surface via Myr47-Bio+SA was far more efficient than that to scavenger receptor class B type 1 (SR-B1). Furthermore, liposomes bound to cell surface NTCP via Myr47-Bio+SA were endocytosed into cells after cells were cultured at 37 °C. However, this endocytosis was suppressed by 4 °C or cytochalasin B treatment. (4) Conclusions: This model system will be useful for not only analyzing HBV entry mechanisms but also screening substances to prevent HBV infection. |
| first_indexed | 2024-03-09T15:45:22Z |
| format | Article |
| id | doaj.art-0812dee0e5694379ae8b03b6f8a1681a |
| institution | Directory Open Access Journal |
| issn | 2076-393X |
| language | English |
| last_indexed | 2024-03-09T15:45:22Z |
| publishDate | 2022-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Vaccines |
| spelling | doaj.art-0812dee0e5694379ae8b03b6f8a1681a2023-11-24T18:31:49ZengMDPI AGVaccines2076-393X2022-11-011012205010.3390/vaccines10122050Specific Binding and Endocytosis of Liposomes to HEK293T Cells via Myrisoylated Pre-S1 Peptide Bound to Sodium Taurocholate Cotransporting PolypeptideShuji Hinuma0Kazuyo Fujita1Shun’ichi Kuroda2Department of Biomolecular Science and Reaction, SANKEN, Osaka University, Mihogaoka 8-1, Osaka 567-0047, JapanDepartment of Biomolecular Science and Reaction, SANKEN, Osaka University, Mihogaoka 8-1, Osaka 567-0047, JapanDepartment of Biomolecular Science and Reaction, SANKEN, Osaka University, Mihogaoka 8-1, Osaka 567-0047, Japan(1) Background: Sodium taurocholate cotransporting polypeptide (NTCP) functions as a key receptor for the hepatitis B virus (HBV) infection. Analyzing HBV and NTCP interaction is an important issue not only for basic research but also for the development of anti-HBV therapeutics. We developed here a novel model system to analyze the interaction of NTCP with liposomes instead of HBV. (2) Methods: Liposomal binding and endocytosis through NTCP in HEK293T cells were achieved by serial treatments of HEL293T cells transiently expressing NTCP-green fluorescence protein (GFP) fusion protein with a synthetic biotinylated pre-S1 peptide (Myr47-Bio) and streptavidin (SA) complex (i.e., Myr47-Bio+SA) followed by biotinylated liposomes. By this procedure, binding of [biotinylated liposomes]-[Myr47-Bio+SA]-[NTCP-GFP] was formed. (3) Results: Using this model system, we found that liposomal binding to NTCP on the cell surface via Myr47-Bio+SA was far more efficient than that to scavenger receptor class B type 1 (SR-B1). Furthermore, liposomes bound to cell surface NTCP via Myr47-Bio+SA were endocytosed into cells after cells were cultured at 37 °C. However, this endocytosis was suppressed by 4 °C or cytochalasin B treatment. (4) Conclusions: This model system will be useful for not only analyzing HBV entry mechanisms but also screening substances to prevent HBV infection.https://www.mdpi.com/2076-393X/10/12/2050hepatitis B virus (HBV)sodium cholate cotransporting peptide (NCP)liposomemyristoylated pre-S1 domain peptidereceptor bindingendocytosis |
| spellingShingle | Shuji Hinuma Kazuyo Fujita Shun’ichi Kuroda Specific Binding and Endocytosis of Liposomes to HEK293T Cells via Myrisoylated Pre-S1 Peptide Bound to Sodium Taurocholate Cotransporting Polypeptide Vaccines hepatitis B virus (HBV) sodium cholate cotransporting peptide (NCP) liposome myristoylated pre-S1 domain peptide receptor binding endocytosis |
| title | Specific Binding and Endocytosis of Liposomes to HEK293T Cells via Myrisoylated Pre-S1 Peptide Bound to Sodium Taurocholate Cotransporting Polypeptide |
| title_full | Specific Binding and Endocytosis of Liposomes to HEK293T Cells via Myrisoylated Pre-S1 Peptide Bound to Sodium Taurocholate Cotransporting Polypeptide |
| title_fullStr | Specific Binding and Endocytosis of Liposomes to HEK293T Cells via Myrisoylated Pre-S1 Peptide Bound to Sodium Taurocholate Cotransporting Polypeptide |
| title_full_unstemmed | Specific Binding and Endocytosis of Liposomes to HEK293T Cells via Myrisoylated Pre-S1 Peptide Bound to Sodium Taurocholate Cotransporting Polypeptide |
| title_short | Specific Binding and Endocytosis of Liposomes to HEK293T Cells via Myrisoylated Pre-S1 Peptide Bound to Sodium Taurocholate Cotransporting Polypeptide |
| title_sort | specific binding and endocytosis of liposomes to hek293t cells via myrisoylated pre s1 peptide bound to sodium taurocholate cotransporting polypeptide |
| topic | hepatitis B virus (HBV) sodium cholate cotransporting peptide (NCP) liposome myristoylated pre-S1 domain peptide receptor binding endocytosis |
| url | https://www.mdpi.com/2076-393X/10/12/2050 |
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