Influence of Subcellular Localization and Functional State on Protein Turnover
Protein homeostasis is an equilibrium of paramount importance that maintains cellular performance by preserving an efficient proteome. This equilibrium avoids the accumulation of potentially toxic proteins, which could lead to cellular stress and death. While the regulators of proteostasis are the m...
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MDPI AG
2021-07-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/10/7/1747 |
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| author | Roya Yousefi Kristina Jevdokimenko Verena Kluever David Pacheu-Grau Eugenio F. Fornasiero |
| author_facet | Roya Yousefi Kristina Jevdokimenko Verena Kluever David Pacheu-Grau Eugenio F. Fornasiero |
| author_sort | Roya Yousefi |
| collection | DOAJ |
| description | Protein homeostasis is an equilibrium of paramount importance that maintains cellular performance by preserving an efficient proteome. This equilibrium avoids the accumulation of potentially toxic proteins, which could lead to cellular stress and death. While the regulators of proteostasis are the machineries controlling protein production, folding and degradation, several other factors can influence this process. Here, we have considered two factors influencing protein turnover: the subcellular localization of a protein and its functional state. For this purpose, we used an imaging approach based on the pulse-labeling of 17 representative SNAP-tag constructs for measuring protein lifetimes. With this approach, we obtained precise measurements of protein turnover rates in several subcellular compartments. We also tested a selection of mutants modulating the function of three extensively studied proteins, the Ca<sup>2+</sup> sensor calmodulin, the small GTPase Rab5a and the brain creatine kinase (CKB). Finally, we followed up on the increased lifetime observed for the constitutively active Rab5a (Q79L), and we found that its stabilization correlates with enlarged endosomes and increased interaction with membranes. Overall, our data reveal that both changes in protein localization and functional state are key modulators of protein turnover, and protein lifetime fluctuations can be considered to infer changes in cellular behavior. |
| first_indexed | 2024-03-10T09:42:43Z |
| format | Article |
| id | doaj.art-07b9ebb2b70141b09dd11adeaab3ddec |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-10T09:42:43Z |
| publishDate | 2021-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-07b9ebb2b70141b09dd11adeaab3ddec2023-11-22T03:29:25ZengMDPI AGCells2073-44092021-07-01107174710.3390/cells10071747Influence of Subcellular Localization and Functional State on Protein TurnoverRoya Yousefi0Kristina Jevdokimenko1Verena Kluever2David Pacheu-Grau3Eugenio F. Fornasiero4Department of Neuro- and Sensory Physiology, University Medical Center Göttingen, 37073 Göttingen, GermanyDepartment of Neuro- and Sensory Physiology, University Medical Center Göttingen, 37073 Göttingen, GermanyDepartment of Neuro- and Sensory Physiology, University Medical Center Göttingen, 37073 Göttingen, GermanyDepartment of Cellular Biochemistry, University Medical Center Göttingen, 37073 Göttingen, GermanyDepartment of Neuro- and Sensory Physiology, University Medical Center Göttingen, 37073 Göttingen, GermanyProtein homeostasis is an equilibrium of paramount importance that maintains cellular performance by preserving an efficient proteome. This equilibrium avoids the accumulation of potentially toxic proteins, which could lead to cellular stress and death. While the regulators of proteostasis are the machineries controlling protein production, folding and degradation, several other factors can influence this process. Here, we have considered two factors influencing protein turnover: the subcellular localization of a protein and its functional state. For this purpose, we used an imaging approach based on the pulse-labeling of 17 representative SNAP-tag constructs for measuring protein lifetimes. With this approach, we obtained precise measurements of protein turnover rates in several subcellular compartments. We also tested a selection of mutants modulating the function of three extensively studied proteins, the Ca<sup>2+</sup> sensor calmodulin, the small GTPase Rab5a and the brain creatine kinase (CKB). Finally, we followed up on the increased lifetime observed for the constitutively active Rab5a (Q79L), and we found that its stabilization correlates with enlarged endosomes and increased interaction with membranes. Overall, our data reveal that both changes in protein localization and functional state are key modulators of protein turnover, and protein lifetime fluctuations can be considered to infer changes in cellular behavior.https://www.mdpi.com/2073-4409/10/7/1747protein stabilityoptical analysis of protein turnoverSNAP-tagpulse-chaseRab5a |
| spellingShingle | Roya Yousefi Kristina Jevdokimenko Verena Kluever David Pacheu-Grau Eugenio F. Fornasiero Influence of Subcellular Localization and Functional State on Protein Turnover Cells protein stability optical analysis of protein turnover SNAP-tag pulse-chase Rab5a |
| title | Influence of Subcellular Localization and Functional State on Protein Turnover |
| title_full | Influence of Subcellular Localization and Functional State on Protein Turnover |
| title_fullStr | Influence of Subcellular Localization and Functional State on Protein Turnover |
| title_full_unstemmed | Influence of Subcellular Localization and Functional State on Protein Turnover |
| title_short | Influence of Subcellular Localization and Functional State on Protein Turnover |
| title_sort | influence of subcellular localization and functional state on protein turnover |
| topic | protein stability optical analysis of protein turnover SNAP-tag pulse-chase Rab5a |
| url | https://www.mdpi.com/2073-4409/10/7/1747 |
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