| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Molecular & Cellular Proteomics 1:376-386, 2002.
© 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
,¶,||,**
Protein Interaction Laboratory, University of Southern Denmark
MDS-Proteomics A/S, Staermosegaardsvej 6, Odense M, DK-5230, Denmark
Quantitative proteomics has traditionally been performed by two-dimensional gel electrophoresis, but recently, mass spectrometric methods based on stable isotope quantitation have shown great promise for the simultaneous and automated identification and quantitation of complex protein mixtures. Here we describe a method, termed SILAC, for stable isotope labeling by amino acids in cell culture, for the in vivo incorporation of specific amino acids into all mammalian proteins. Mammalian cell lines are grown in media lacking a standard essential amino acid but supplemented with a non-radioactive, isotopically labeled form of that amino acid, in this case deuterated leucine (Leu-d3). We find that growth of cells maintained in these media is no different from growth in normal media as evidenced by cell morphology, doubling time, and ability to differentiate. Complete incorporation of Leu-d3 occurred after five doublings in the cell lines and proteins studied. Protein populations from experimental and control samples are mixed directly after harvesting, and mass spectrometric identification is straightforward as every leucine-containing peptide incorporates either all normal leucine or all Leu-d3. We have applied this technique to the relative quantitation of changes in protein expression during the process of muscle cell differentiation. Proteins that were found to be up-regulated during this process include glyceraldehyde-3-phosphate dehydrogenase, fibronectin, and pyruvate kinase M2. SILAC is a simple, inexpensive, and accurate procedure that can be used as a quantitative proteomic approach in any cell culture system.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  M. Mann and N. L. Kelleher Mass Spectrometry Special Feature: Precision proteomics: The case for high resolution and high mass accuracy PNAS, November 25, 2008; 105(47): 18132 - 18138. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. Trinkle-Mulcahy, S. Boulon, Y. W. Lam, R. Urcia, F.-M. Boisvert, F. Vandermoere, N. A. Morrice, S. Swift, U. Rothbauer, H. Leonhardt, et al. Identifying specific protein interaction partners using quantitative mass spectrometry and bead proteomes J. Cell Biol., October 20, 2008; 183(2): 223 - 239. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. Song, W. Kong, N. Weatherspoon, G. Qin, W. Tyler, J. Turk, R. Curtiss III, and Y. Shi Modulation of the Regulatory Activity of Bacterial Two-component Systems by SlyA J. Biol. Chem., October 17, 2008; 283(42): 28158 - 28168. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. P. Mirza and M. Olivier Methods and approaches for the comprehensive characterization and quantification of cellular proteomes using mass spectrometry Physiol Genomics, October 8, 2008; 33(1): 3 - 11. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. W. Deutsch, H. Lam, and R. Aebersold Data analysis and bioinformatics tools for tandem mass spectrometry in proteomics Physiol Genomics, October 8, 2008; 33(1): 18 - 25. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Liao, S. K. Park, T. Xu, P. Vanderklish, and J. R. Yates III Quantitative proteomic analysis of primary neurons reveals diverse changes in synaptic protein content in fmr1 knockout mice PNAS, October 7, 2008; 105(40): 15281 - 15286. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Skiba, T. C. Mettenleiter, and A. Karger Quantitative Whole-Cell Proteome Analysis of Pseudorabies Virus-Infected Cells J. Virol., October 1, 2008; 82(19): 9689 - 9699. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 V. Zabrouskov, Y. Ge, J. Schwartz, and J. W. Walker Unraveling Molecular Complexity of Phosphorylated Human Cardiac Troponin I by Top Down Electron Capture Dissociation/Electron Transfer Dissociation Mass Spectrometry Mol. Cell. Proteomics, October 1, 2008; 7(10): 1838 - 1849. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C.-L. Han, C.-W. Chien, W.-C. Chen, Y.-R. Chen, C.-P. Wu, H. Li, and Y.-J. Chen A Multiplexed Quantitative Strategy for Membrane Proteomics: Opportunities for Mining Therapeutic Targets for Autosomal Dominant Polycystic Kidney Disease Mol. Cell. Proteomics, October 1, 2008; 7(10): 1983 - 1997. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Doucet, G. S. Butler, D. Rodriguez, A. Prudova, and C. M. Overall Metadegradomics: Toward in Vivo Quantitative Degradomics of Proteolytic Post-translational Modifications of the Cancer Proteome Mol. Cell. Proteomics, October 1, 2008; 7(10): 1925 - 1951. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
U. Kruse, M. Bantscheff, G. Drewes, and C. Hopf Chemical and Pathway Proteomics: Powerful Tools for Oncology Drug Discovery and Personalized Health Care Mol. Cell. Proteomics, October 1, 2008; 7(10): 1887 - 1901. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Ota, K. Kito, S. Okada, and T. Ito A proteomic screen reveals the mitochondrial outer membrane protein Mdm34p as an essential target of the F-box protein Mdm30p Genes Cells, October 1, 2008; 13(10): 1075 - 1085. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Lebreton, J.-C. Rousselle, P. Lenormand, A. Namane, A. Jacquier, M. Fromont-Racine, and C. Saveanu 60S ribosomal subunit assembly dynamics defined by semi-quantitative mass spectrometry of purified complexes Nucleic Acids Res., September 1, 2008; 36(15): 4988 - 4999. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. C. Bendall, C. Hughes, M. H. Stewart, B. Doble, M. Bhatia, and G. A. Lajoie Prevention of Amino Acid Conversion in SILAC Experiments with Embryonic Stem Cells Mol. Cell. Proteomics, September 1, 2008; 7(9): 1587 - 1597. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Bantscheff, M. Boesche, D. Eberhard, T. Matthieson, G. Sweetman, and B. Kuster Robust and Sensitive iTRAQ Quantification on an LTQ Orbitrap Mass Spectrometer Mol. Cell. Proteomics, September 1, 2008; 7(9): 1702 - 1713. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Mertins, H. C. Eberl, J. Renkawitz, J. V. Olsen, M. L. Tremblay, M. Mann, A. Ullrich, and H. Daub Investigation of Protein-tyrosine Phosphatase 1B Function by Quantitative Proteomics Mol. Cell. Proteomics, September 1, 2008; 7(9): 1763 - 1777. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Raijmakers, C. R. Berkers, A. de Jong, H. Ovaa, A. J. R. Heck, and S. Mohammed Automated Online Sequential Isotope Labeling for Protein Quantitation Applied to Proteasome Tissue-specific Diversity Mol. Cell. Proteomics, September 1, 2008; 7(9): 1755 - 1762. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Luo, Y. Li, J.-J. Mu, J. Zhang, T. Tonaka, Y. Hamamori, S. Y. Jung, Y. Wang, and J. Qin Regulation of Intra-S Phase Checkpoint by Ionizing Radiation (IR)-dependent and IR-independent Phosphorylation of SMC3 J. Biol. Chem., July 11, 2008; 283(28): 19176 - 19183. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. P. Albuquerque, M. B. Smolka, S. H. Payne, V. Bafna, J. Eng, and H. Zhou A Multidimensional Chromatography Technology for In-depth Phosphoproteome Analysis Mol. Cell. Proteomics, July 1, 2008; 7(7): 1389 - 1396. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. S. Spellman, K. Deinhardt, C. C. Darie, M. V. Chao, and T. A. Neubert Stable Isotopic Labeling by Amino Acids in Cultured Primary Neurons: Application to Brain-derived Neurotrophic Factor-dependent Phosphotyrosine-associated Signaling Mol. Cell. Proteomics, June 1, 2008; 7(6): 1067 - 1076. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Mousson, A. Kolkman, W. W. M. P. Pijnappel, H. Th. M. Timmers, and A. J. R. Heck Quantitative Proteomics Reveals Regulation of Dynamic Components within TATA-binding Protein (TBP) Transcription Complexes Mol. Cell. Proteomics, May 1, 2008; 7(5): 845 - 852. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Colzani, F. Schutz, A. Potts, P. Waridel, and M. Quadroni Relative Protein Quantification by Isobaric SILAC with Immonium Ion Splitting (ISIS) Mol. Cell. Proteomics, May 1, 2008; 7(5): 927 - 937. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. M.R. Lacerda, L. Xin, I. Rogers, and K. F. Reardon Analysis of iTRAQ data using Mascot and Peaks quantification algorithms Brief Funct Genomic Proteomic, April 4, 2008; (2008) eln017v1. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Graumann, N. C. Hubner, J. B. Kim, K. Ko, M. Moser, C. Kumar, J. Cox, H. Scholer, and M. Mann Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) and Proteome Quantitation of Mouse Embryonic Stem Cells to a Depth of 5,111 Proteins Mol. Cell. Proteomics, April 1, 2008; 7(4): 672 - 683. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Panchaud, J. Hansson, M. Affolter, R. Bel Rhlid, S. Piu, P. Moreillon, and M. Kussmann ANIBAL, Stable Isotope-based Quantitative Proteomics by Aniline and Benzoic Acid Labeling of Amino and Carboxylic Groups Mol. Cell. Proteomics, April 1, 2008; 7(4): 800 - 812. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. W. H. Wong, M. J. Sullivan, and G. Cagney Computational methods for the comparative quantification of proteins in label-free LCn-MS experiments Brief Bioinform, March 1, 2008; 9(2): 156 - 165. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. A. Stead, N. W. Paton, P. Missier, S. M. Embury, C. Hedeler, B. Jin, A. J. P. Brown, and A. Preece Information quality in proteomics Brief Bioinform, March 1, 2008; 9(2): 174 - 188. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. G. Xiao, M. Garg, S. Lim, D. Wong, V. L. Go, and W.-N. P. Lee Determination of protein synthesis in vivo using labeling from deuterated water and analysis of MALDI-TOF spectrum J Appl Physiol, March 1, 2008; 104(3): 828 - 836. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Villanueva, A. Nazarian, K. Lawlor, S. S. Yi, R. J. Robbins, and P. Tempst A Sequence-specific Exopeptidase Activity Test (SSEAT) for "Functional" Biomarker Discovery Mol. Cell. Proteomics, March 1, 2008; 7(3): 509 - 518. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Kruger, I. Kratchmarova, B. Blagoev, Y.-H. Tseng, C. R. Kahn, and M. Mann Dissection of the insulin signaling pathway via quantitative phosphoproteomics PNAS, February 19, 2008; 105(7): 2451 - 2456. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Nanavati, M. Gucek, J. L. S. Milne, S. Subramaniam, and S. P. Markey Stoichiometry and Absolute Quantification of Proteins with Mass Spectrometry Using Fluorescent and Isotope-labeled Concatenated Peptide Standards Mol. Cell. Proteomics, February 1, 2008; 7(2): 442 - 447. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Guo, J. Villen, J. Kornhauser, K. A. Lee, M. P. Stokes, K. Rikova, A. Possemato, J. Nardone, G. Innocenti, R. Wetzel, et al. Signaling networks assembled by oncogenic EGFR and c-Met PNAS, January 15, 2008; 105(2): 692 - 697. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Diella, C. M. Gould, C. Chica, A. Via, and T. J. Gibson Phospho.ELM: a database of phosphorylation sites update 2008 Nucleic Acids Res., January 11, 2008; 36(suppl_1): D240 - D244. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. J. Pesavento, H. Yang, N. L. Kelleher, and C. A. Mizzen Certain and Progressive Methylation of Histone H4 at Lysine 20 during the Cell Cycle Mol. Cell. Biol., January 1, 2008; 28(1): 468 - 486. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Hebeler, S. Oeljeklaus, K. A. Reidegeld, M. Eisenacher, C. Stephan, B. Sitek, K. Stuhler, H. E. Meyer, M. J. G. Sturre, P. P. Dijkwel, et al. Study of Early Leaf Senescence in Arabidopsis thaliana by Quantitative Proteomics Using Reciprocal 14N/15N Labeling and Difference Gel Electrophoresis Mol. Cell. Proteomics, January 1, 2008; 7(1): 108 - 120. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. L. Miller, S. Hanke, A. M. Hinsby, C. Friis, S. Brunak, M. Mann, and N. Blom Motif Decomposition of the Phosphotyrosine Proteome Reveals a New N-terminal Binding Motif for SHIP2 Mol. Cell. Proteomics, January 1, 2008; 7(1): 181 - 192. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J.-W. Yang, H. Vacher, K.-S. Park, E. Clark, and J. S. Trimmer Trafficking-dependent phosphorylation of Kv1.2 regulates voltage-gated potassium channel cell surface expression PNAS, December 11, 2007; 104(50): 20055 - 20060. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. P. Stokes, J. Rush, J. MacNeill, J. M. Ren, K. Sprott, J. Nardone, V. Yang, S. A. Beausoleil, S. P. Gygi, M. Livingstone, et al. Profiling of UV-induced ATM/ATR signaling pathways PNAS, December 11, 2007; 104(50): 19855 - 19860. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Pisitkun, J. D. Hoffert, M.-J. Yu, and M. A. Knepper Tandem Mass Spectrometry in Physiology Physiology, December 1, 2007; 22(6): 390 - 400. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Keshishian, T. Addona, M. Burgess, E. Kuhn, and S. A. Carr Quantitative, Multiplexed Assays for Low Abundance Proteins in Plasma by Targeted Mass Spectrometry and Stable Isotope Dilution Mol. Cell. Proteomics, December 1, 2007; 6(12): 2212 - 2229. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Bonenfant, H. Towbin, M. Coulot, P. Schindler, D. R. Mueller, and J. van Oostrum Analysis of Dynamic Changes in Post-translational Modifications of Human Histones during Cell Cycle by Mass Spectrometry Mol. Cell. Proteomics, November 1, 2007; 6(11): 1917 - 1932. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L.-Y. Tang, N. Deng, L.-S. Wang, J. Dai, Z.-L. Wang, X.-S. Jiang, S.-J. Li, L. Li, Q.-H. Sheng, D.-Q. Wu, et al. Quantitative Phosphoproteome Profiling of Wnt3a-mediated Signaling Network: Indicating the Involvement of Ribonucleoside-diphosphate Reductase M2 Subunit Phosphorylation at Residue Serine 20 in Canonical Wnt Signal Transduction Mol. Cell. Proteomics, November 1, 2007; 6(11): 1952 - 1967. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. A. Everley, C. A. Gartner, W. Haas, A. Saghatelian, J. E. Elias, B. F. Cravatt, B. R. Zetter, and S. P. Gygi Assessing Enzyme Activities Using Stable Isotope Labeling and Mass Spectrometry Mol. Cell. Proteomics, October 1, 2007; 6(10): 1771 - 1777. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B.-J. M. Webb-Robertson and W. R. Cannon Current trends in computational inference from mass spectrometry-based proteomics Brief Bioinform, September 1, 2007; 8(5): 304 - 317. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. Coute, C. Hernandez, R. D. Appel, J.-C. Sanchez, and A. Margolles Labeling of Bifidobacterium longum Cells with 13C-Substituted Leucine for Quantitative Proteomic Analyses Appl. Envir. Microbiol., September 1, 2007; 73(17): 5653 - 5656. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Bouyssie, A. G. de Peredo, E. Mouton, R. Albigot, L. Roussel, N. Ortega, C. Cayrol, O. Burlet-Schiltz, J.-P. Girard, and B. Monsarrat Mascot File Parsing and Quantification (MFPaQ), a New Software to Parse, Validate, and Quantify Proteomics Data Generated by ICAT and SILAC Mass Spectrometric Analyses: Application To the Proteomics Study of Membrane Proteins from Primary Human Endothelial Cells Mol. Cell. Proteomics, September 1, 2007; 6(9): 1621 - 1637. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. R. Cutillas and B. Vanhaesebroeck Quantitative Profile of Five Murine Core Proteomes Using Label-free Functional Proteomics Mol. Cell. Proteomics, September 1, 2007; 6(9): 1560 - 1573. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. B. McClatchy, L. Liao, S. K. Park, J. D. Venable, and J. R. Yates Quantification of the synaptosomal proteome of the rat cerebellum during post-natal development Genome Res., September 1, 2007; 17(9): 1378 - 1388. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Rivers, D. M. Simpson, D. H. L. Robertson, S. J. Gaskell, and R. J. Beynon Absolute Multiplexed Quantitative Analysis of Protein Expression during Muscle Development Using QconCAT Mol. Cell. Proteomics, August 1, 2007; 6(8): 1416 - 1427. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. McIlwain, D. Page, E. L. Huttlin, and M. R. Sussman Using dynamic programming to create isotopic distribution maps from mass spectra Bioinformatics, July 1, 2007; 23(13): i328 - i336. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. J. Benschop, S. Mohammed, M. O'Flaherty, A. J. R. Heck, M. Slijper, and F. L. H. Menke Quantitative Phosphoproteomics of Early Elicitor Signaling in Arabidopsis Mol. Cell. Proteomics, July 1, 2007; 6(7): 1198 - 1214. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. L. Maxwell and L. Frappier Viral Proteomics Microbiol. Mol. Biol. Rev., June 1, 2007; 71(2): 398 - 411. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. S. Lane, Y. Wang, R. Betts, W. J. Griffiths, and L. H. Patterson Comparative Cytochrome P450 Proteomics in the Livers of Immunodeficient Mice Using 18O Stable Isotope Labeling Mol. Cell. Proteomics, June 1, 2007; 6(6): 953 - 962. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Jin, G. J. Li, J. Davis, D. Zhu, Y. Wang, C. Pan, and J. Zhang Identification of Novel Proteins Associated with Both {alpha}-Synuclein and DJ-1 Mol. Cell. Proteomics, May 1, 2007; 6(5): 845 - 859. [Abstract] [Full Text] [PDF]
|
