The DOI matches its Crossref record; 1 of the 1 references it cites were found in a registry or an index.
A Direct Nanoflow Liquid Chromatography−Tandem Mass Spectrometry System for Interaction Proteomics
Abstract
One of the strategies of functional proteomics, research aiming to discover gene function at the protein level, is the comprehensive analysis of protein-protein interactions related to the functional linkage among proteins and analysis of functional cellular machinery to better understand the basis of cell functions. Here, we describe the direct nanoflow LC (DNLC) system, which is equipped with a fritless high-resolution electrospray interface column packed with 1-microm reversed-phase (RP) beads and a novel splitless nanoflow gradient elution system to operate the column. Using RP-DNLC at an extremely slow flow rate, <50 nL/min, combined with data-dependent collision-induced dissociation tandem MS (MS/MS) and computer-assisted retrieval of spectra, we identified approximately 100 protein components in a biological complex such as a premature mammalian ribosome pull-down from cultured cells when we used an epitope-tagged protein as bait. Because this analysis is most sensitive, requires approximately 0.2 microg of total protein, and is a fully automated 1-h process, we anticipated that it should be an excellent tool for analyzing a limited amount of functional multi-protein complexes in cells.
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Cites
A comprehensive analysis of protein–protein interactions in Saccharomyces cerevisiae
Uetz, P., Giot, L., Cagney, G., Mansfield, T. A., Judson, R. S., Knight, J. R., Lockshon, D., Narayan, V., Srinivasan, M., Pochart, P., et al. (2000) · Nature · doi:10.1038/35001009 · Cited by 4,706
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Nutrient-dependent mTORC1 Association with the ULK1–Atg13–FIP200 Complex Required for Autophagy
Hosokawa, N., Hara, T., Kaizuka, T., Kishi, C., Takamura, A., Miura, Y., Iemura, S.-I., Natsume, T., Takehana, K., Yamada, N., et al. (2009) · Molecular Biology of the Cell · doi:10.1091/mbc.e08-12-1248 · Cited by 2,076
Self-citationFIP200, a ULK-interacting protein, is required for autophagosome formation in mammalian cells
Hara, T., Takamura, A., Kishi, C., Iemura, S.-I., Natsume, T., Guan, J.-L., & Mizushima, N. (2008) · The Journal of Cell Biology · doi:10.1083/jcb.200712064 · Cited by 904
Self-citationMizushima, N., Kuma, A., Kobayashi, Y., Yamamoto, A., Matsubae, M., Takao, T., Natsume, T., Ohsumi, Y., & Yoshimori, T. (2003) · Journal of Cell Science · doi:10.1242/jcs.00381 · Cited by 791
Self-citationThe HOPS complex mediates autophagosome–lysosome fusion through interaction with syntaxin 17
Jiang, P., Nishimura, T., Sakamaki, Y., Itakura, E., Hatta, T., Natsume, T., & Mizushima, N. (2014) · Molecular Biology of the Cell · doi:10.1091/mbc.e13-08-0447 · Cited by 501
Self-citationMoriguchi, T., Urushiyama, S., Hisamoto, N., Iemura, S.-I., Uchida, S., Natsume, T., Matsumoto, K., & Shibuya, H. (2005) · Journal of Biological Chemistry · doi:10.1074/jbc.m510042200 · Cited by 477
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Metadata
- Authors
- Natsume, T., Yamauchi, Y., Nakayama, H., Shinkawa, T., Yanagida, M., Takahashi, N., & Isobe, T.
- Venue
- Analytical Chemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 2002-08-10
- Volume
- 74
- Issue
- 18
- Pages
- 4725-4733
- ISSN
- 0003-2700 (ISSN-L)
- 1520-6882
- OpenAlex
- W1967404904
- Cited by
- 193
- References
- 1
- FWCI
- 6.28
- Type
- journal article
- Language
- en
- Open access
- closed
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