Changes in Protein Expression in Warmed Human Lens Epithelium Cells Using Shotgun Proteomics
<i>Background and Objectives</i>: In previous studies, we reported that the assessment of the cumulative thermal dose in the crystalline lens, conducted through computational modeling utilizing a supercomputer and the biothermal transport equation, exhibited a significant association wit...
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MDPI AG
2025-02-01
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| author | Hiroko Otake Tetsushi Yamamoto Naoki Yamamoto Yosuke Nakazawa Yoshiki Miyata Atsushi Taga Hiroshi Sasaki Noriaki Nagai |
| author_facet | Hiroko Otake Tetsushi Yamamoto Naoki Yamamoto Yosuke Nakazawa Yoshiki Miyata Atsushi Taga Hiroshi Sasaki Noriaki Nagai |
| author_sort | Hiroko Otake |
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| description | <i>Background and Objectives</i>: In previous studies, we reported that the assessment of the cumulative thermal dose in the crystalline lens, conducted through computational modeling utilizing a supercomputer and the biothermal transport equation, exhibited a significant association with the incidence of nuclear cataracts. In this study, we have investigated the types of proteins that expressed underlying 35.0 °C (normal-temp) and 37.5 °C (warming-temp) by using the shotgun liquid chromatography (LC) with tandem mass spectrometry (MS/MS)-based global proteomic approach. <i>Materials and Methods</i>: We have discussed the changes in protein expression in warmed iHLEC-NY2 cells using Gene Ontology analysis and a label-free semiquantitative method based on spectral counting. <i>Results</i>: In iHLEC-NY2, 615 proteins were detected, including 307 (49.9%) present in both lenses cultured at normal-temp and warming-temp, 130 (21.1%) unique to the lens cultured at normal-temp, and 178 (29.0%) unique to the lens cultured at warming-temp. Furthermore, LC–MS/MS analysis showed that warming decreased the expression of actin, alpha cardiac muscle 1, actin-related protein 2, putative tubulin-like protein alpha-4B, ubiquitin carboxyl-terminal hydrolase 17-like protein 1, ubiquitin-ribosomal protein eL40 fusion protein, ribosome biogenesis protein BMS1 homolog, histone H2B type 1-M, and histone H2A.J. in iHLEC-NY2. <i>Conclusions</i>: The decreases in the specific protein levels of actin, tubulin, ubiquitin, ribosomes, and histones may be related to cataract development under warming conditions. This investigation could provide a critical framework for understanding the correlation between temperature dynamics and the development of nuclear cataracts. |
| format | Article |
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| institution | DOAJ |
| issn | 1010-660X 1648-9144 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | MDPI AG |
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| series | Medicina |
| spelling | doaj-art-040302e2fcea48cb94bcab35752b3d6d2025-08-20T02:44:46ZengMDPI AGMedicina1010-660X1648-91442025-02-0161228610.3390/medicina61020286Changes in Protein Expression in Warmed Human Lens Epithelium Cells Using Shotgun ProteomicsHiroko Otake0Tetsushi Yamamoto1Naoki Yamamoto2Yosuke Nakazawa3Yoshiki Miyata4Atsushi Taga5Hiroshi Sasaki6Noriaki Nagai7Faculty of Pharmacy, Kindai University, Osaka 577-8502, JapanFaculty of Pharmacy, Kindai University, Osaka 577-8502, JapanResearch Promotion Headquarters, Fujita Health University, Toyoake 470-1192, JapanFaculty of Pharmacy, Keio University, 1-5-30 Shibakoen, Tokyo 105-8512, JapanFaculty of Pharmacy, Teikyo University, 2-11-1 Kaga, Tokyo 173-8606, JapanFaculty of Pharmacy, Kindai University, Osaka 577-8502, JapanDepartment of Ophthalmology, Kanazawa Medical University, Uchinada 920-0293, JapanFaculty of Pharmacy, Kindai University, Osaka 577-8502, Japan<i>Background and Objectives</i>: In previous studies, we reported that the assessment of the cumulative thermal dose in the crystalline lens, conducted through computational modeling utilizing a supercomputer and the biothermal transport equation, exhibited a significant association with the incidence of nuclear cataracts. In this study, we have investigated the types of proteins that expressed underlying 35.0 °C (normal-temp) and 37.5 °C (warming-temp) by using the shotgun liquid chromatography (LC) with tandem mass spectrometry (MS/MS)-based global proteomic approach. <i>Materials and Methods</i>: We have discussed the changes in protein expression in warmed iHLEC-NY2 cells using Gene Ontology analysis and a label-free semiquantitative method based on spectral counting. <i>Results</i>: In iHLEC-NY2, 615 proteins were detected, including 307 (49.9%) present in both lenses cultured at normal-temp and warming-temp, 130 (21.1%) unique to the lens cultured at normal-temp, and 178 (29.0%) unique to the lens cultured at warming-temp. Furthermore, LC–MS/MS analysis showed that warming decreased the expression of actin, alpha cardiac muscle 1, actin-related protein 2, putative tubulin-like protein alpha-4B, ubiquitin carboxyl-terminal hydrolase 17-like protein 1, ubiquitin-ribosomal protein eL40 fusion protein, ribosome biogenesis protein BMS1 homolog, histone H2B type 1-M, and histone H2A.J. in iHLEC-NY2. <i>Conclusions</i>: The decreases in the specific protein levels of actin, tubulin, ubiquitin, ribosomes, and histones may be related to cataract development under warming conditions. This investigation could provide a critical framework for understanding the correlation between temperature dynamics and the development of nuclear cataracts.https://www.mdpi.com/1648-9144/61/2/286shotgun proteomicslenstemperaturecataractiHLEC-NY2 |
| spellingShingle | Hiroko Otake Tetsushi Yamamoto Naoki Yamamoto Yosuke Nakazawa Yoshiki Miyata Atsushi Taga Hiroshi Sasaki Noriaki Nagai Changes in Protein Expression in Warmed Human Lens Epithelium Cells Using Shotgun Proteomics Medicina shotgun proteomics lens temperature cataract iHLEC-NY2 |
| title | Changes in Protein Expression in Warmed Human Lens Epithelium Cells Using Shotgun Proteomics |
| title_full | Changes in Protein Expression in Warmed Human Lens Epithelium Cells Using Shotgun Proteomics |
| title_fullStr | Changes in Protein Expression in Warmed Human Lens Epithelium Cells Using Shotgun Proteomics |
| title_full_unstemmed | Changes in Protein Expression in Warmed Human Lens Epithelium Cells Using Shotgun Proteomics |
| title_short | Changes in Protein Expression in Warmed Human Lens Epithelium Cells Using Shotgun Proteomics |
| title_sort | changes in protein expression in warmed human lens epithelium cells using shotgun proteomics |
| topic | shotgun proteomics lens temperature cataract iHLEC-NY2 |
| url | https://www.mdpi.com/1648-9144/61/2/286 |
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