Temperature sensitive Mutant Proteome Profiling: a novel tool for the characterization of the global impact of missense mutations on the proteome
Open accessPreprint — published as 10.1074/jbc.ra120.014576 · Paper page
Abstract
Temperature sensitive (TS) mutants have been foundational in the characterization of essential genes. However, a high-throughput workflow for characterization of biophysical changes in TS mutants is lacking. Te mperature sensitive M utant P roteome P rofiling (TeMPP) is a novel application of mass spectrometry (MS) based thermal proteome profiling (TPP) to characterize effects of missense mutations on protein stability and PPIs. This study characterizes missense mutations in two different subunits of the 26S proteasome on the thermal stability of the proteome at large, revealing distinct mechanistic details that were not obtained using only steady-state transcriptome and proteome analyses. TeMPP is a precise approach to measure changes in missense mutant containing proteomes without the requirement for large amounts of starting material, specific antibodies against proteins of interest, and/or genetic manipulation of the biological system. Overall, TeMPP provides unique mechanistic insights into missense mutation dysfunction and connection of genotype to phenotype in a rapid, non-biased fashion.
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NF-κB Signaling Is Regulated by Fucosylation in Metastatic Breast Cancer Cells
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In silico λ-dynamics predicts protein binding specificities to modified RNAs
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Self-citation
- Authors
- Peck Justice, S. A., Qi, G., Wijeratne, H. R. S., Victorino, J. F., Simpson, E. R., Wijeratne, A. B., & Mosley, A. L.
- Affiliations
- Indiana University Indianapolis (US)
- Corresponding author
- Wijeratne, Aruna B.
- Mosley, Amber L.
- Venue
- Journal of Biological Chemistry
- Publisher
- openRxiv
- Published
- 2019-12-30
- Volume
- 295
- Issue
- 48
- Pages
- 16219-16238
- ISSN
- 0021-9258 (ISSN-L)
- PMC
- PMC7705321
- OpenAlex
- W3082270748
- Cited by
- 28
- References
- 156
- Citations by year
- 2025: 4 · 2024: 6 · 2023: 3 · 2022: 6 · 2021: 7 · 2020: 2
- FWCI
- 1.72
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Biological Chemistry (publishedVersion, cc-by)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- IUScholarWorks (Indiana University) (submittedVersion, cc-by)
- Funders
- Eli Lilly and Company
- Indiana Clinical and Translational Sciences Institute (UL1TR002529)
- Lilly Endowment
- Ralph W. and Grace M. Showalter Research Trust Fund
- Thermo Fisher Scientific
- National Institutes of Health (UL1TR002529, HL007910)
- School of Medicine, Indiana University (UL1TR002529)
- National Heart, Lung, and Blood Institute (HL007910, T32HL007910)
- National Institute of General Medical Sciences (R01GM099714)
- National Center for Advancing Translational Sciences
- Topics
- Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Heat shock proteins research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ubiquitin and proteasome pathways
- Keywords
- missense mutations, temperature sensitivity, 26S proteasome, proteasome subunits, protein stability, genotype-phenotype correlation
- MeSH terms
- RNA-Seq, Saccharomyces cerevisiae, Temperature, Mutation, Missense, Proteome, Saccharomyces cerevisiae Proteins, Proteasome Endopeptidase Complex
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