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Global Analysis of Protein Activities Using Proteome Chips
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To facilitate studies of the yeast proteome, we cloned 5800 open reading frames and overexpressed and purified their corresponding proteins. The proteins were printed onto slides at high spatial density to form a yeast proteome microarray and screened for their ability to interact with proteins and phospholipids. We identified many new calmodulin- and phospholipid-interacting proteins; a common potential binding motif was identified for many of the calmodulin-binding proteins. Thus, microarrays of an entire eukaryotic proteome can be prepared and screened for diverse biochemical activities. The microarrays can also be used to screen protein-drug interactions and to detect posttranslational modifications.
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“Prior to the development of NAPPA, protein microarrays were not widely adopted as a result of difficulties in generating purified proteins to spot onto the arrays by linkage chemistry [105–108].”
From Proteomic approaches to uncovering virus–host protein interactions during the progression of viral infection (2016) · cites this paper as [108]
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- For details of 96-well format protein purification protocol a full list of results from all the experiments and the design of the positive identification algorithms please visit our public Web site () and supplementary material at Science Online (www.sciencemag.org/cgi/content/full/1062191/DC1).
- 10.1016/S1567-1356(01)00005-8 · DOI
- Positives were identified using a combination of the GenePix software which computes a local intensity background for each spot and a series of algorithms we developed. Details can be found at and at www.sciencemag.org/cgi/content/full/1062191/DC1.
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Metadata
- Authors
- Zhu, H., Bilgin, M., Bangham, R., Hall, D., Casamayor, A., Bertone, P., Lan, N., Jansen, R., Bidlingmaier, S., Houfek, T., Mitchell, T., Miller, P., Dean, R. A., Gerstein, M., & Snyder, M.
- Affiliations
- Department of Molecular, Cellular, and Developmental Biology and
- Yale University (US)
- North Carolina State University (US)
- Corresponding author
- Snyder, Michael
- ORCID
- Venue
- Science
- Published
- 2001-09-14
- Volume
- 293
- Issue
- 5537
- Pages
- 2101-2105
- ISSN
- 0036-8075 (ISSN-L)
- 1095-9203
- OpenAlex
- W4233751197
- Cited by
- 1,884
- References
- 37
- Citations by year
- 2026: 9 · 2025: 16 · 2024: 20 · 2023: 23 · 2022: 37 · 2021: 35 · 2020: 34 · 2019: 33 · 2018: 39 · 2017: 49 · 2016: 56 · 2015: 62 · 2014: 61 · 2013: 60 · 2012: 97
- FWCI
- 47.46
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Science (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Advanced Biosensing Techniques and Applications
- Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Fungal and yeast genetics research
- Keywords
- proteome microarray, yeast proteome, protein-protein interaction, phospholipid binding, calmodulin-binding proteins, calmodulin binding motif, drug-protein interactions, post-translational modification
- MeSH terms
- Amino Acid Sequence, Calmodulin, Calmodulin-Binding Proteins, Cell Membrane, Cloning, Molecular, Fungal Proteins, Glucose, Liposomes, Membrane Proteins, Molecular Sequence Data, Phosphatidylcholines, Phosphatidylinositols, Phospholipids, Protein Binding, Recombinant Fusion Proteins, Saccharomyces cerevisiae, Signal Transduction, Open Reading Frames, Peptide Library, Streptavidin, Proteome, Amino Acid Motifs
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