Industrial-scale proteomics: from liters of plasma to chemically synthesized proteins

Article Article scientifique Article Industrial-scale proteomics: from liters of plasma to chemically synthesized proteins eng 2004 author Rose Keith 21876 author Bougueleret Lydie 639268 author Baussant Thierry author Böhm Günter author Botti Paolo 135016 author Colinge Jacques 29836 author Cusin Isabelle 22966 author Gaertner Hubert François 84280 author Gleizes Anne 651869 author Heller Manfred 133419 author Jimenez Silvia 793858 author Johnson Andrew author Kussmann Martin author Menin Laure author Menzel Christoph author Ranno Frederic author Rodriguez-Tomé Patricia author Rogers John author Saudrais Cedric author Villain Matteo author Wetmore Diana author Bairoch Amos Marc 0000-0003-2826-6444 21964 author Hochstrasser Denise 526168 2125-2150 10.1002/pmic.200300718 15221774 1615-9853 Human blood plasma is a useful source of proteins associated with both health and disease. Analysis of human blood plasma is a challenge due to the large number of peptides and proteins present and the very wide range of concentrations. In order to identify as many proteins as possible for subsequent comparative studies, we developed an industrial-scale (2.5 liter) approach involving sample pooling for the analysis of smaller proteins (M(r) generally < ca. 40 000 and some fragments of very large proteins). Plasma from healthy males was depleted of abundant proteins (albumin and IgG), then smaller proteins and polypeptides were separated into 12 960 fractions by chromatographic techniques. Analysis of proteins and polypeptides was performed by mass spectrometry prior to and after enzymatic digestion. Thousands of peptide identifications were made, permitting the identification of 502 different proteins and polypeptides from a single pool, 405 of which are listed here. The numbers refer to chromatographically separable polypeptide entities present prior to digestion. Combining results from studies with other plasma pools we have identified over 700 different proteins and polypeptides in plasma. Relatively low abundance proteins such as leptin and ghrelin and peptides such as bradykinin, all invisible to two-dimensional gel technology, were clearly identified. Proteins of interest were synthesized by chemical methods for bioassays. We believe that this is the first time that the small proteins in human blood plasma have been separated and analyzed so extensively. Amino Acid Sequence Blood Chemical Analysis/methods Blood Proteins/metabolism Chromatography Chromatography, Gel Chromatography, High Pressure Liquid Chromatography, Ion Exchange Computational Biology Databases as Topic Electrophoresis, Gel, Two-Dimensional/methods Humans Immunoglobulin G/chemistry Mass Spectrometry Molecular Sequence Data Peptides/chemistry Plasma/metabolism Proteome Proteomics/methods Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Subcellular Fractions Time Factors Trypsin/pharmacology Dewey 576 136 Département de biologie structurale et bioinformatique 21E false 283ea908-4a0f-403e-afca-454333963c49 Swiss-Prot Research Group 405b9ec3-ed88-49e8-9c17-4f861cf66d82 Swiss Institute of Bioinformatics (SIB) 57 Calipho 80 211 Auto-immunité, inflammation et athérosclérose 270 Proteomics 4 7 e09b4b97-086b-4609-94e8-39edcc522978 Article (Published version) Restricted pub_137_15221774.pdf 266759 application/pdf restrictedAccess Faculté de médecine / Section de médecine fondamentale / Département de biologie structurale et bioinformatique

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Industrial-scale proteomics: from liters of plasma to chemically synthesized proteins	Proteomics			2004	637	0

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Hochstrasser, Denise