Journal ArticleDOI
In-gel digestion for mass spectrometric characterization of proteins and proteomes
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TLDR
This protocol is for the in-gel digestion of both silver and Coomassie-stained protein spots or bands and can be followed by MALDI-MS or LC-MS/MS analysis to identify proteins at sensitivities better than a few femtomoles of protein starting material.Abstract:
In-gel digestion of proteins isolated by gel electrophoresis is a cornerstone of mass spectrometry (MS)-driven proteomics. The 10-year-old recipe by Shevchenko et al. has been optimized to increase the speed and sensitivity of analysis. The protocol is for the in-gel digestion of both silver and Coomassie-stained protein spots or bands and can be followed by MALDI-MS or LC-MS/MS analysis to identify proteins at sensitivities better than a few femtomoles of protein starting material.read more
Citations
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Targeting the scaffolding role of LSD1 (KDM1A) poises acute myeloid leukemia cells for retinoic acid–induced differentiation
Roberto Ravasio,Elena Ceccacci,Luciano Nicosia,Amir Hosseini,Pier Luigi Rossi,Iros Barozzi,Lorenzo Fornasari,Roberto Dal Zuffo,Sergio Valente,Rossella Fioravanti,Ciro Mercurio,Mario Varasi,Andrea Mattevi,Antonello Mai,Giulio Pavesi,Tiziana Bonaldi,Saverio Minucci,Saverio Minucci +17 more
TL;DR: The histone demethylase LSD1 is deregulated in several tumors, including leukemias, providing the rationale for the clinical use of LSD1 inhibitors, and integration of proteomic/epigenomic/mutational studies showed that LSD1 inhibitor alter the recruitment of LSD 1-containing complexes to chromatin, inhibiting the interaction between LSD1 and the transcription factor GFI1.
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Tick-host conflict: Immunoglobulin E antibodies to tick proteins in patients with anaphylaxis to tick bite
Lourdes Mateos-Hernández,Margarita Villar,Angel Moral,Carmen Garcia Rodríguez,Teresa Alfaya Arias,Verónica de la Osa,Francisco Feo Brito,Isabel G. Fernández de Mera,Pilar Alberdi,Francisco Ruiz-Fons,Alejandro Cabezas-Cruz,Agustín Estrada-Peña,José de la Fuente +12 more
TL;DR: A comparative proteomics approach was used to characterize tick proteins producing an IgE antibody response in a healthy individual with record oftick bites, and two patients with anaphylactic reactions to Rhipicephalus bursa or Hyalomma marginatum tick bites, which supported a patient-specific IgE antibodies response to tick species responsible for theAnaphylaxis to tick bite.
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Amniotic fluid stem cell exosomes: Therapeutic perspective
Francesca Beretti,Manuela Zavatti,Francesca Casciaro,Giuseppina Comitini,Fabrizia Franchi,Veronica Barbieri,Giovanni Battista La Sala,Tullia Maraldi +7 more
TL;DR: Evidence is presented that the principal effect of AFSC conditioned media is the induction of apoptosis in lymphocytes, whereas exposure to AFSC-derived exosomes decreases the lymphocyte's proliferation, supporting the hypothesis that the entire secretome of stem cells differently affects immune-response.
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A Proteomic Survey of Nonribosomal Peptide and Polyketide Biosynthesis in Actinobacteria
Yunqiu Chen,Ioanna Ntai,Kou San Ju,Michelle Unger,Leonid Zamdborg,Sarah J. Robinson,James R. Doroghazi,David P. Labeda,William W. Metcalf,Neil L. Kelleher,Neil L. Kelleher +10 more
TL;DR: The feasibility of a proteomics-based strategy for use in screening for NRP/PK production in actinomycetes versus the bacilli (2-4 Mbp genomes) used previously is demonstrated.
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Ubiquitinated proteins in exosomes secreted by myeloid-derived suppressor cells.
TL;DR: Evidence at the molecular level is provided that ubiquitinated proteins are present in exosomes shed by myeloid-derived suppressor cells (MDSC) and spans a range of protein sizes and favors basic pI values and hydrophobicity.
References
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Journal ArticleDOI
Mass Spectrometric Sequencing of Proteins from Silver-Stained Polyacrylamide Gels
TL;DR: Silver staining allows a substantial shortening of sample preparation time and may, therefore, be preferable over Coomassie staining, and this work removes a major obstacle to the low-level sequence analysis of proteins separated on polyacrylamide gels.
Journal ArticleDOI
Mass spectrometry-based proteomics
Ruedi Aebersold,Matthias Mann +1 more
TL;DR: The ability of mass spectrometry to identify and, increasingly, to precisely quantify thousands of proteins from complex samples can be expected to impact broadly on biology and medicine.
Journal ArticleDOI
Stable isotope labeling by amino acids in cell culture, SILAC, as a simple and accurate approach to expression proteomics.
Shao En Ong,Blagoy Blagoev,Irina Kratchmarova,Dan B. Kristensen,Hanno Steen,Akhilesh Pandey,Matthias Mann +6 more
TL;DR: SILAC is a simple, inexpensive, and accurate procedure that can be used as a quantitative proteomic approach in any cell culture system and is applied to the relative quantitation of changes in protein expression during the process of muscle cell differentiation.
Journal ArticleDOI
Stop and Go extraction tips for matrix-assisted laser desorption/ionization, nanoelectrospray, and lc/ms sample pretreatment in proteomics
TL;DR: A novel procedure in which a very small disk of beads embedded in a Teflon meshwork is placed as a microcolumn into pipet tips, finding that the Stage system is well-suited as a universal sample preparation system for proteomics.
Journal ArticleDOI
Mass spectrometry-based proteomics turns quantitative.
Shao En Ong,Matthias Mann +1 more
TL;DR: Two recently developed methodologies offer the opportunity to obtain quantitative proteomic information by comparing the signals from the same peptide under different conditions, and stable isotope labels facilitates direct quantification from the mass spectra.
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