| High hydrostatic pressure inhibits the biosynthesis of eukaryotic elongation factor-2. | |
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MedLine Citation:
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PMID: 15534876 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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High continuous hydrostatic pressure is known to inhibit the total cellular protein synthesis. In this study, our goal was to identify pressure-regulated proteins by using two dimensional gel electrophoresis and mass spectrometry. This analysis showed that under 30 MPa continuous hydrostatic pressure the biosynthesis of eukaryotic elongation factor-2 (eEF-2) was inhibited both in HeLa carcinoma and T/C28a4 chondrocytic cell lines. Western blot analysis of HeLa cells revealed that the cellular protein level of eEF-2 decreased by 40%-50% within 12 h of the pressure treatment. However, the steady-state mRNA level of eEF-2 was not affected by the pressure. Cycloheximide addition after 4 h-pressure treatment suggested that the half-life of eEF-2 protein was shorter in pressurized cells. eEF-2 is responsible for the translocation of ribosome along the specific mRNA during translation, and its phosphorylation prevents the ribosomal translocation. Therefore, increased phosphorylation of eEF-2 was considered as one mechanism that could explain the reduced level of protein synthesis in pressurized HeLa cell cultures. However, Western blot analysis with an antibody recognizing the Thr56-phosphorylated form of eEF-2 showed that phosphorylation of eEF-2 was not elevated in pressurized samples. In conclusion, the inhibition of protein synthesis under high pressure occurs independent of the phosphorylation of eEF-2. However, this inhibition may result from the decrease of cellular eEF-2 protein. |
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Authors:
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Mika A Elo; Hannu M Karjalainen; Reijo K Sironen; Leena Valmu; Nicholas T Redpath; Gareth J Browne; Nisse Kalkkinen; Heikki J Helminen; Mikko J Lammi |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Journal of cellular biochemistry Volume: 94 ISSN: 0730-2312 ISO Abbreviation: J. Cell. Biochem. Publication Date: 2005 Feb |
Date Detail:
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Created Date: 2005-01-26 Completed Date: 2005-08-09 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 8205768 Medline TA: J Cell Biochem Country: United States |
Other Details:
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Languages: eng Pagination: 497-507 Citation Subset: IM |
Copyright Information:
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Copyright 2004 Wiley-Liss, Inc. |
Affiliation:
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Department of Anatomy, University of Kuopio, 70211 Kuopio, Finland. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Amino Acid Sequence Base Sequence Blotting, Northern Blotting, Western DNA Primers Electrophoresis, Gel, Two-Dimensional Eukaryotic Cells / metabolism Hela Cells Humans Molecular Sequence Data Peptide Elongation Factor 2 / biosynthesis*, chemistry, metabolism Phosphorylation Pressure Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization |
| Chemical | |
Reg. No./Substance:
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0/DNA Primers; 0/Peptide Elongation Factor 2 |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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