Activated Protein Kinase C (PKC) Is Persistently Trafficked with Epidermal Growth Factor (EGF) Receptor
Protein kinase Cs (PKCs) are activated by lipids in the plasma membrane and bind to a scaffold assembled on the epidermal growth factor (EGF) receptor (EGFR). Understanding how this complex is routed is important, because this determines whether EGFR is degraded, terminating signaling. Here, cells w...
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| Format: | Article |
| Language: | English |
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MDPI AG
2020-09-01
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| Series: | Biomolecules |
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| Online Access: | https://www.mdpi.com/2218-273X/10/9/1288 |
| _version_ | 1797554320393109504 |
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| author | Carol A. Heckman Tania Biswas Douglas M. Dimick Marilyn L. Cayer |
| author_facet | Carol A. Heckman Tania Biswas Douglas M. Dimick Marilyn L. Cayer |
| author_sort | Carol A. Heckman |
| collection | DOAJ |
| description | Protein kinase Cs (PKCs) are activated by lipids in the plasma membrane and bind to a scaffold assembled on the epidermal growth factor (EGF) receptor (EGFR). Understanding how this complex is routed is important, because this determines whether EGFR is degraded, terminating signaling. Here, cells were preincubated in EGF-tagged gold nanoparticles, then allowed to internalize them in the presence or absence of a phorbol ester PKC activator. PKC colocalized with EGF-tagged nanoparticles within 5 min and migrated with EGFR-bearing vesicles into the cell. Two conformations of PKC-epsilon were distinguished by different primary antibodies. One, thought to be enzymatically active, was on endosomes and displayed a binding site for antibody RR (R&D). The other, recognized by Genetex green (GG), was soluble, on actin-rich structures, and loosely bound to vesicles. During a 15-min chase, EGF-tagged nanoparticles entered large, perinuclear structures. In phorbol ester-treated cells, vesicles bearing EGF-tagged nanoparticles tended to enter this endocytic recycling compartment (ERC) without the GG form. The correlation coefficient between the GG (inactive) and RR conformations on vesicles was also lower. Thus, active PKC has a Charon-like function, ferrying vesicles to the ERC, and inactivation counteracts this function. The advantage conferred on cells by aggregating vesicles in the ERC is unclear. |
| first_indexed | 2024-03-10T16:30:20Z |
| format | Article |
| id | doaj.art-0bbf39c0e9ef478eae0374f6abcee715 |
| institution | Directory Open Access Journal |
| issn | 2218-273X |
| language | English |
| last_indexed | 2024-03-10T16:30:20Z |
| publishDate | 2020-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomolecules |
| spelling | doaj.art-0bbf39c0e9ef478eae0374f6abcee7152023-11-20T12:52:07ZengMDPI AGBiomolecules2218-273X2020-09-01109128810.3390/biom10091288Activated Protein Kinase C (PKC) Is Persistently Trafficked with Epidermal Growth Factor (EGF) ReceptorCarol A. Heckman0Tania Biswas1Douglas M. Dimick2Marilyn L. Cayer3Department of Biological Sciences, 217 Life Science Building, Bowling Green State University, Bowling Green, OH 43403, USADepartment of Biological Sciences, 217 Life Science Building, Bowling Green State University, Bowling Green, OH 43403, USADepartment of Physics & Astronomy, 104 Overman Hall, Bowling Green State University, Bowling Green, OH 43403, USACenter for Microscopy & Microanalysis, 217 Life Science Building, Bowling Green State University, Bowling Green, OH 43403, USAProtein kinase Cs (PKCs) are activated by lipids in the plasma membrane and bind to a scaffold assembled on the epidermal growth factor (EGF) receptor (EGFR). Understanding how this complex is routed is important, because this determines whether EGFR is degraded, terminating signaling. Here, cells were preincubated in EGF-tagged gold nanoparticles, then allowed to internalize them in the presence or absence of a phorbol ester PKC activator. PKC colocalized with EGF-tagged nanoparticles within 5 min and migrated with EGFR-bearing vesicles into the cell. Two conformations of PKC-epsilon were distinguished by different primary antibodies. One, thought to be enzymatically active, was on endosomes and displayed a binding site for antibody RR (R&D). The other, recognized by Genetex green (GG), was soluble, on actin-rich structures, and loosely bound to vesicles. During a 15-min chase, EGF-tagged nanoparticles entered large, perinuclear structures. In phorbol ester-treated cells, vesicles bearing EGF-tagged nanoparticles tended to enter this endocytic recycling compartment (ERC) without the GG form. The correlation coefficient between the GG (inactive) and RR conformations on vesicles was also lower. Thus, active PKC has a Charon-like function, ferrying vesicles to the ERC, and inactivation counteracts this function. The advantage conferred on cells by aggregating vesicles in the ERC is unclear.https://www.mdpi.com/2218-273X/10/9/1288receptor tyrosine kinaseslow endocytic recyclingannexincortactinvesicle trafficking |
| spellingShingle | Carol A. Heckman Tania Biswas Douglas M. Dimick Marilyn L. Cayer Activated Protein Kinase C (PKC) Is Persistently Trafficked with Epidermal Growth Factor (EGF) Receptor Biomolecules receptor tyrosine kinase slow endocytic recycling annexin cortactin vesicle trafficking |
| title | Activated Protein Kinase C (PKC) Is Persistently Trafficked with Epidermal Growth Factor (EGF) Receptor |
| title_full | Activated Protein Kinase C (PKC) Is Persistently Trafficked with Epidermal Growth Factor (EGF) Receptor |
| title_fullStr | Activated Protein Kinase C (PKC) Is Persistently Trafficked with Epidermal Growth Factor (EGF) Receptor |
| title_full_unstemmed | Activated Protein Kinase C (PKC) Is Persistently Trafficked with Epidermal Growth Factor (EGF) Receptor |
| title_short | Activated Protein Kinase C (PKC) Is Persistently Trafficked with Epidermal Growth Factor (EGF) Receptor |
| title_sort | activated protein kinase c pkc is persistently trafficked with epidermal growth factor egf receptor |
| topic | receptor tyrosine kinase slow endocytic recycling annexin cortactin vesicle trafficking |
| url | https://www.mdpi.com/2218-273X/10/9/1288 |
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