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1.
PLoS One ; 15(1): e0226770, 2020.
Article in English | MEDLINE | ID: mdl-31945070

ABSTRACT

Despite the increasing importance of non-targeted metabolomics to answer various life science questions, extracting biochemically relevant information from metabolomics spectral data is still an incompletely solved problem. Most computational tools to identify tandem mass spectra focus on a limited set of molecules of interest. However, such tools are typically constrained by the availability of reference spectra or molecular databases, limiting their applicability of generating structural hypotheses for unknown metabolites. In contrast, recent advances in the field illustrate the possibility to expose the underlying biochemistry without relying on metabolite identification, in particular via substructure prediction. We describe an automated method for substructure recommendation motivated by association rule mining. Our framework captures potential relationships between spectral features and substructures learned from public spectral libraries. These associations are used to recommend substructures for any unknown mass spectrum. Our method does not require any predefined metabolite candidates, and therefore it can be used for the hypothesis generation or partial identification of unknown unknowns. The method is called MESSAR (MEtabolite SubStructure Auto-Recommender) and is implemented in a free online web service available at messar.biodatamining.be.


Subject(s)
Biological Products/analysis , Databases, Factual , Metabolome , Pharmaceutical Preparations/analysis , Tandem Mass Spectrometry/methods , Automation , Humans
2.
Anal Chim Acta ; 1044: 181-190, 2018 Dec 31.
Article in English | MEDLINE | ID: mdl-30442400

ABSTRACT

Antibody-based pharmaceuticals often encompass a complex structural heterogeneity requiring enhanced analytical methods for reliable characterization of variants and degradation products. We have explored the capabilities of low-flow sheathless capillary electrophoresis-mass spectrometry (CE-MS) for the high-resolution and sensitive profiling of antibody therapeutics. Near-zero electroosmotic flow was achieved by employing a novel neutral capillary coating that also prevents protein adsorption. CE-MS analysis of intact model proteins using an acidic background electrolyte demonstrated satisfactory performance, with overall migration-time RSDs below 2.2% from three different capillaries tested. For system evaluation, three nanobody preparations, including mono- and bivalent forms, and three monoclonal antibodies (mAbs) were analyzed. Intact nanobodies were resolved from their degradation products, which could be assigned to deamidated, cleaved, and truncated forms at the C-terminal tag. Excellent resolution of isomeric deamidated products was obtained. The mAbs were analyzed intact and after digestion by the endoproteinase IdeS (middle-up approach). CE-MS of intact mAbs provided resolution of clipped species (e.g. light chain and light chain-heavy chain fragments) from the native protein. Moreover, glycoforms containing sialic acids were resolved from their non-sialylated counterparts. For IdeS-digested, F (ab)2 and Fc/2 portions where efficiently resolved for the three mAbs. Whereas the migration time of the Fc/2 fragments was fairly similar, the migration time of the F (ab)2 part was strongly varied among the mAbs. For all mAbs, separation of Fc/2 charge variants - including sialylated glycoforms and other post-translational modifications, such as loss of C-terminal lysine or asparagine deamidation - was achieved. This allowed a detailed and reliable assessment of the Fc/2 heterogeneity (18-33 proteoforms) of the three analyzed mAbs.


Subject(s)
Antibodies, Monoclonal/analysis , Antibodies, Monoclonal/chemistry , Electrophoresis, Capillary/methods , Mass Spectrometry/methods , Antibodies, Monoclonal/pharmacology , Antibodies, Monoclonal/therapeutic use , Immunoglobulin Fc Fragments/chemistry , Sialic Acids/chemistry
3.
Mass Spectrom Rev ; 37(5): 607-629, 2018 09.
Article in English | MEDLINE | ID: mdl-29120505

ABSTRACT

The identification of unknown molecules has been one of the cornerstone applications of mass spectrometry for decades. This tutorial reviews the basics of the interpretation of electrospray ionization-based MS and MS/MS spectra in order to identify small-molecule analytes (typically below 2000 Da). Most of what is discussed in this tutorial also applies to other atmospheric pressure ionization methods like atmospheric pressure chemical/photoionization. We focus primarily on the fundamental steps of MS-based structural elucidation of individual unknown compounds, rather than describing strategies for large-scale identification in complex samples. We critically discuss topics like the detection of protonated and deprotonated ions ([M + H]+ and [M - H]- ) as well as other adduct ions, the determination of the molecular formula, and provide some basic rules on the interpretation of product ion spectra. Our tutorial focuses primarily on the fundamental steps of MS-based structural elucidation of individual unknown compounds (eg, contaminants in chemical production, pharmacological alteration of drugs), rather than describing strategies for large-scale identification in complex samples. This tutorial also discusses strategies to obtain useful orthogonal information (UV/Vis, H/D exchange, chemical derivatization, etc) and offers an overview of the different informatics tools and approaches that can be used for structural elucidation of small molecules. It is primarily intended for beginning mass spectrometrists and researchers from other mass spectrometry sub-disciplines that want to get acquainted with structural elucidation are interested in some practical tips and tricks.

4.
Anal Chim Acta ; 982: 122-130, 2017 Aug 22.
Article in English | MEDLINE | ID: mdl-28734351

ABSTRACT

Site-specific mapping of multiple deamidations in peptides is a challenging analytical task. In this work, capillary electrophoresis-tandem mass spectrometry (CE-MS/MS) is presented as a high-resolution tool for the detailed characterization of these subtle modifications in peptides. The 4.5-kDa peptide drug TRI-1144, which contains five closely-positioned potential deamidation sites, was selected as model compound. TRI-1144 was exposed to acidic conditions and/or elevated temperatures for 1-14 h. Stressed samples were analyzed using a background electrolyte (BGE) of 150 mM ammonium formate (pH 6.0) in combination with a capillary coated with a bilayer of Polybrene-dextran sulfate. Separation of deamidated and deacetylated TRI-1144 species, including several positional isomers, was greatly enhanced by adding up to 40 vol% of acetonitrile-isopropanol (87.5:12.5, v/v) to the BGE, allowing reliable determination of the number of deamidations/deacetylations per degradation product. Collision-induced dissociation MS/MS was conducted on the separated peptide components in order to reveal the exact position of deamidation on the peptide chain. Obtained fragment ions showed overlapping isotopic distributions in their MS/MS spectra resulting from the comigration of different isomeric deamidated species. Comparison of theoretical and measured isotope distributions for specific y ions of peptide fragments yielded the identity and relative abundance of isomeric deamidated products. The developed CE-MS/MS methodology was used for the highly selective evaluation of TRI-1144 stability under different stress conditions, providing detailed qualitative and semi-quantitative degradation maps of the peptide drug.


Subject(s)
Electrophoresis, Capillary , Peptides/chemistry , Tandem Mass Spectrometry
5.
J Pharm Sci ; 106(5): 1310-1316, 2017 05.
Article in English | MEDLINE | ID: mdl-28108379

ABSTRACT

Forced degradation studies are an important tool for a systematic assessment of decomposition pathways and identification of reactive sites in active pharmaceutical ingredients (APIs). Two methodologies have been combined in order to provide a deeper understanding of singlet oxygen-related degradation pathways of APIs under light irradiation. First, we report that a "dark" singlet oxygen test enables the investigation of drug reactivity toward singlet oxygen independently of photolytic irradiation processes. Second, the photosensitizing properties of the API producing the singlet oxygen was proven and quantified by spin trapping and electron paramagnetic resonance analysis. A combination of these techniques is an interesting addition to the forced degradation portfolio as it can be used for (1) revealing unexpected degradation pathways of APIs due to singlet oxygen, (2) clarifying photolytic drug-drug interactions in fixed-dose combinations, and (3) synthesizing larger quantities of hardly accessible oxidative drug degradants.


Subject(s)
Pharmaceutical Preparations/chemistry , Photolysis , Singlet Oxygen/chemistry , Spin Trapping/methods , Electron Spin Resonance Spectroscopy/methods , Light/adverse effects , Oxidants, Photochemical/chemistry , Oxidants, Photochemical/metabolism , Oxidation-Reduction , Pharmaceutical Preparations/metabolism , Singlet Oxygen/metabolism
6.
Int J Parasitol ; 41(10): 1079-92, 2011 Aug 15.
Article in English | MEDLINE | ID: mdl-21723868

ABSTRACT

Giardia lamblia is a flagellated protozoan parasite and a major cause of diarrhoea in humans. Its microtubular cytoskeleton mediates trophozoite motility, attachment and cytokinesis, and is characterised by an attachment disk and eight flagella that are each nucleated in a basal body. To date, only 10 giardial basal body proteins have been identified, including universal signalling proteins that are important for regulating mitosis or differentiation. In this study, we have exploited bioinformatics and proteomic approaches to identify new Giardia basal body proteins and confocal microscopy to confirm their localisation in interphase trophozoites. This approach identified 75 homologs of conserved basal body proteins in the genome including 65 not previously known to be associated with Giardia basal bodies. Thirteen proteins were confirmed to co-localise with centrin to the Giardia basal bodies. We also demonstrate that most basal body proteins localise to additional cytoskeletal structures in interphase trophozoites. This might help to explain the roles of the four pairs of flagella and Giardia-specific organelles in motility and differentiation. A deeper understanding of the composition of the Giardia basal bodies will contribute insights into the complex signalling pathways that regulate its unique cytoskeleton and the biological divergence of these conserved organelles.


Subject(s)
Genome, Protozoan , Giardia lamblia/chemistry , Giardia lamblia/genetics , Organelles/chemistry , Organelles/genetics , Proteome/analysis , Protozoan Proteins/analysis , Computational Biology , Genes, Protozoan , Microscopy, Confocal
7.
Proc Natl Acad Sci U S A ; 106(40): 17019-24, 2009 Oct 06.
Article in English | MEDLINE | ID: mdl-19805154

ABSTRACT

Plasma cells daily secrete their own mass in antibodies, which fold and assemble in the endoplasmic reticulum (ER). To reach these levels, cells require pERp1, a novel lymphocyte-specific small ER-resident protein, which attains expression levels as high as BiP when B cells differentiate into plasma cells. Although pERp1 has no homology with known ER proteins, it does contain a CXXC motif typical for oxidoreductases. In steady state, the CXXC cysteines are locked by two parallel disulfide bonds with a downstream C(X)(6)C motif, and pERp1 displays only modest oxidoreductase activity. pERp1 emerged as a dedicated folding factor for IgM, associating with both heavy and light chains and promoting assembly and secretion of mature IgM.


Subject(s)
Endoplasmic Reticulum/metabolism , Immunoglobulin M/metabolism , Molecular Chaperones/metabolism , Plasma Cells/metabolism , Amino Acid Sequence , Animals , B-Lymphocytes/metabolism , B-Lymphocytes/ultrastructure , Cell Differentiation , Cell Line, Tumor , Electrophoresis, Gel, Two-Dimensional , Endoplasmic Reticulum Chaperone BiP , HeLa Cells , Heat-Shock Proteins/genetics , Heat-Shock Proteins/metabolism , Humans , Immunoblotting , Mass Spectrometry , Mice , Microscopy, Fluorescence , Microscopy, Immunoelectron , Molecular Chaperones/genetics , Oxidoreductases/metabolism , Plasma Cells/cytology , RNA Interference , Sulfhydryl Compounds/metabolism
8.
Mol Biol Cell ; 20(4): 1150-66, 2009 Feb.
Article in English | MEDLINE | ID: mdl-19109428

ABSTRACT

Centrioles are intriguing cylindrical organelles composed of triplet microtubules. Proteomic data suggest that a large number of proteins besides tubulin are necessary for the formation and maintenance of a centriole's complex structure. Expansion of the preexisting centriole proteome from the green alga Chlamydomonas reinhardtii revealed additional human disease genes, emphasizing the significance of centrioles in normal human tissue homeostasis. We found that two classes of ciliary disease genes were highly represented among the basal body proteome: cystic kidney disease (especially nephronophthisis) syndromes, including Meckel/Joubert-like and oral-facial-digital syndrome, caused by mutations in CEP290, MKS1, OFD1, and AHI1/Jouberin proteins and cone-rod dystrophy syndrome genes, including UNC-119/HRG4, NPHP4, and RPGR1. We further characterized proteome of the centriole (POC) 1, a highly abundant WD40 domain-containing centriole protein. We found that POC1 is recruited to nascent procentrioles and localizes in a highly asymmetrical pattern in mature centrioles corresponding to sites of basal-body fiber attachment. Knockdown of POC1 in human cells caused a reduction in centriole duplication, whereas overexpression caused the appearance of elongated centriole-like structures. Together, these data suggest that POC1 is involved in early steps of centriole duplication as well as in the later steps of centriole length control.


Subject(s)
Algal Proteins/chemistry , Algal Proteins/metabolism , Centrioles/metabolism , Chlamydomonas reinhardtii/cytology , Chlamydomonas reinhardtii/metabolism , Conserved Sequence , Proteome/metabolism , Amino Acid Sequence , Animals , Axoneme/metabolism , Axoneme/ultrastructure , Centrioles/ultrastructure , Chlamydomonas reinhardtii/ultrastructure , Disease/genetics , Fluorescent Antibody Technique , Humans , Microtubules/metabolism , Microtubules/ultrastructure , Molecular Sequence Data , Mutation , Protein Structure, Tertiary , Protein Transport , RNA Interference , Repetitive Sequences, Amino Acid
9.
J Cell Biol ; 178(6): 905-12, 2007 Sep 10.
Article in English | MEDLINE | ID: mdl-17785518

ABSTRACT

Basal bodies organize the nine doublet microtubules found in cilia. Cilia are required for a variety of cellular functions, including motility and sensing stimuli. Understanding this biochemically complex organelle requires an inventory of the molecular components and the contribution each makes to the overall structure. We define a basal body proteome and determine the specific localization of basal body components in the ciliated protozoan Tetrahymena thermophila. Using a biochemical, bioinformatic, and genetic approach, we identify 97 known and candidate basal body proteins. 24 novel T. thermophila basal body proteins were identified, 19 of which were localized to the ultrastructural level, as seen by immunoelectron microscopy. Importantly, we find proteins from several structural domains within the basal body, allowing us to reveal how each component contributes to the overall organization. Thus, we present a high resolution localization map of basal body structure highlighting important new components for future functional studies.


Subject(s)
Centrioles/metabolism , Protozoan Proteins/metabolism , Tetrahymena thermophila/metabolism , Animals , Centrioles/ultrastructure , Cilia/metabolism , Cilia/ultrastructure , Microscopy, Electron, Transmission , Microtubules/metabolism , Microtubules/ultrastructure , Proteome/metabolism , Tetrahymena thermophila/ultrastructure
10.
Curr Biol ; 15(12): 1090-8, 2005 Jun 21.
Article in English | MEDLINE | ID: mdl-15964273

ABSTRACT

BACKGROUND: The centriole is one of the most enigmatic organelles in the cell. Centrioles are cylindrical, microtubule-based barrels found in the core of the centrosome. Centrioles also act as basal bodies during interphase to nucleate the assembly of cilia and flagella. There are currently only a handful of known centriole proteins. RESULTS: We used mass-spectrometry-based MudPIT (multidimensional protein identification technology) to identify the protein composition of basal bodies (centrioles) isolated from the green alga Chlamydomonas reinhardtii. This analysis detected the majority of known centriole proteins, including centrin, epsilon tubulin, and the cartwheel protein BLD10p. By combining proteomic data with information about gene expression and comparative genomics, we identified 45 cross-validated centriole candidate proteins in two classes. Members of the first class of proteins (BUG1-BUG27) are encoded by genes whose expression correlates with flagellar assembly and which therefore may play a role in ciliogenesis-related functions of basal bodies. Members of the second class (POC1-POC18) are implicated by comparative-genomics and -proteomics studies to be conserved components of the centriole. We confirmed centriolar localization for the human homologs of four candidate proteins. Three of the cross-validated centriole candidate proteins are encoded by orthologs of genes (OFD1, NPHP-4, and PACRG) implicated in mammalian ciliary function and disease, suggesting that oral-facial-digital syndrome and nephronophthisis may involve a dysfunction of centrioles and/or basal bodies. CONCLUSIONS: By analyzing isolated Chlamydomonas basal bodies, we have been able to obtain the first reported proteomic analysis of the centriole.


Subject(s)
Centrioles/genetics , Chlamydomonas/genetics , Cilia/genetics , Proteomics/methods , Protozoan Proteins/genetics , Amino Acid Sequence , Animals , Cilia/pathology , Conserved Sequence , Humans , Kidney Diseases/genetics , Microfilament Proteins , Molecular Chaperones , Nuclear Proteins/genetics , Orofaciodigital Syndromes/genetics , Proteins/genetics , Protozoan Proteins/analysis , Reproducibility of Results
11.
Mol Cell Proteomics ; 4(9): 1297-310, 2005 Sep.
Article in English | MEDLINE | ID: mdl-15961381

ABSTRACT

B cells play an essential role in the immune response. Upon activation they may differentiate into plasma cells that secrete specific antibodies against potentially pathogenic non-self antigens. To identify the cellular proteins that are important for efficient production of these antibodies we set out to study the B cell differentiation process at the proteome level. We performed an in-depth proteomic study to quantify dynamic relative protein expression patterns of several hundreds of proteins at five consecutive time points after lipopolysaccharide-induced activation of B lymphocytes. The proteome analysis was performed using a combination of stable isotope labeling using [13C6]leucine added to the murine B cell cultures, one-dimensional gel electrophoresis, and LC-MS/MS. In this study we identified 1,001 B cell proteins. We were able to quantify the expression levels of a quarter of all identified proteins (i.e. 234) at each of the five different time points. Nearly all proteins revealed changes in expression patterns. The quantitative dataset was further analyzed using an unbiased clustering method. Based on their expression profiles, we grouped the entire set of 234 quantified proteins into a limited number of 12 distinct clusters. Functionally related proteins showed a strong correlation in their temporal expression profiles. The quality of the quantitative data allowed us to even identify subclusters within functionally related classes of proteins such as in the endoplasmic reticulum proteins that are involved in antibody production.


Subject(s)
B-Lymphocytes/physiology , Cell Differentiation , Chromatography, Liquid , Electrophoresis , Mass Spectrometry , Proteome/metabolism , Proteomics , Amino Acids/metabolism , Animals , Carbon Isotopes , Cell Line, Tumor , Isotope Labeling , Leucine/metabolism , Lipopolysaccharides/pharmacology , Lymphocyte Activation/drug effects , Mice , Models, Biological , Reference Standards
12.
Proteomics ; 4(8): 2397-407, 2004 Aug.
Article in English | MEDLINE | ID: mdl-15274135

ABSTRACT

Tyrosyl radicals cross-linked to protein tyrosine residues (tyrosylated proteins) represent hallmarks of neutrophil-mediated injury at the inflammatory locus. Yet the proteins targeted by tyrosyl radicals in an intact cellular system remain to be elucidated. Here, we show that tyrosyl radicals generated by human neutrophils after activation by phorbol 12-myristate 13-acetate (PMA), interferon-gamma (IFN-gamma) or TNF-alpha could act in an autocrine manner by cross-linking to endogenous proteins. We have identified the tyrosylated proteins by using a membrane-impermeable tyrosine analogue, tyramine coupled to fluorescein (TyrFluo), in combination with proteomics techniques. Confocal microscopy images indicated that initially the tyrosylated proteins were localized in patches at the cell surface to become internalized subsequently. In the neutrophil membrane-associated proteome, lactoferrin was the prime target of tyrosylation. Out of three isoforms identified, an 80 kDa neutral isoform was tyrosylated more extensively than the 85 kD basic isoform, particularly after PMA activation. Although all three stimuli induced tyrosylation of the filamentous component vimentin, additional tyrosylated vimentin fragments were detected after IFN-gamma- and TNF-alpha-stimulation. Moreover, upon activation the bulk of vimentin behaved as a dimer (M(r) 120 kDa) being slightly tyrosylated, yet phosphorylated at Thr-425 possibly as a requirement for its externalization. Unexpectedly, bovine catalase added to end tyrosyl radicals formation was detected as a highly tyrosylated neutrophil-associated protein. A moderate stimulus-dependent tyrosylation of ATP synthase-beta, alpha-enolase, glyceraldehyde 3-phosphate dehydrogenase, cytokeratin-10, filamin-A, and annexin-I was also observed. When the membrane-permeable probe (acetylTyrFluo) was used, protein tyrosylation was not observed indicating that the intracellular proteins were well protected against oxidative attack. This study shows that human neutrophils can modulate their proteome via a tyrosine oxidation pathway induced by pro-inflammatory mediators.


Subject(s)
Fluorescent Dyes/chemistry , Free Radicals/chemistry , Neutrophil Activation , Neutrophils/chemistry , Proteins/analysis , Proteome/analysis , Tyrosine , Animals , Blotting, Western , Cattle , Electrophoresis, Gel, Two-Dimensional , Humans , Interferon-gamma/pharmacology , Mass Spectrometry , Molecular Sequence Data , Neutrophils/drug effects , Peptide Mapping , Tetradecanoylphorbol Acetate/pharmacology , Tumor Necrosis Factor-alpha/pharmacology , Tyrosine/analogs & derivatives , Tyrosine/chemistry
13.
Proteomics ; 3(10): 2008-18, 2003 Oct.
Article in English | MEDLINE | ID: mdl-14625863

ABSTRACT

A mouse model for nonatopic asthma was employed to study the alterations of the lung proteome to gain insight into the underlying molecular mechanisms of disease pathophysiology post-challenge. Lung samples from asthmatic and control mice were used to generate 24 high quality two-dimensional electrophoresis gels wherein 2115 proteins were examined for disease relevance. In total, 23 proteins were significantly up- or down-regulated following hapten-challenge of dinitro-fluorobenzene-hypersensitive mice. Twenty proteins were identified by mass spectrometry, of which 18 could be linked to asthma related symptoms, such as stress and inflammation, lung detoxification, plasma exudation and/or tissue remodeling. As such, proteomics was clearly vindicated as a means of studying this complex disease phenomenon. The proteins found in this study may not necessarily play a role in the immunological mechanisms and/or pathophysiology of asthma development. However, they may prove useful as surrogate biomarkers for quantitatively monitoring disease state progression or response to therapy. The mathematics of achieving statistical confidence from low numbers of gel replicates containing large numbers of independent variables stress the need for high numbers of replicates to better sample the population of proteins revealed by two-dimensional gel electrophoresis.


Subject(s)
Asthma/metabolism , Hypersensitivity, Delayed/metabolism , Lung/chemistry , Proteome/analysis , Actins/analysis , Alcohol Oxidoreductases/analysis , Animals , Annexin A6/analysis , Asthma/immunology , Asthma/physiopathology , Benzenesulfonates/immunology , Benzenesulfonates/pharmacology , Data Interpretation, Statistical , Dinitrofluorobenzene/immunology , Dinitrofluorobenzene/pharmacology , Disease Models, Animal , Electrophoresis, Gel, Two-Dimensional , Gas Chromatography-Mass Spectrometry , Gelsolin/analysis , Glutathione Transferase/analysis , Heat-Shock Proteins/analysis , Hypersensitivity, Delayed/immunology , Hypersensitivity, Delayed/physiopathology , Isoelectric Focusing , Lung/drug effects , Mice , Mice, Inbred BALB C , Microfilament Proteins , Nerve Tissue Proteins/analysis , Proteins/analysis , Proteome/metabolism , Proteomics/methods , Serum Albumin/analysis , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Talin/analysis , Vinculin/analysis
14.
J Chromatogr A ; 1000(1-2): 589-608, 2003 Jun 06.
Article in English | MEDLINE | ID: mdl-12877191

ABSTRACT

Conventional proteomics makes use of two-dimensional gel electrophoresis followed by mass spectrometric analysis of typtic fragments derived from in-gel digestion of proteins. Although being a very strong technique capable of separating and visualizing hundreds of proteins, 2D-gel electrophoresis has some well-documented disadvantages as well. More recently, liquid chromatographic-(tandem) mass spectrometric techniques have been developed to overcome some of the shortcomings of 2D-gel electrophoresis. In this review we have described several recent applications of liquid chromatography-(tandem) mass spectrometry in the field of proteomics and especially in the field of membrane proteomics, quantitative proteomics and in the analysis of post-translational modifications.


Subject(s)
Chromatography, Liquid/methods , Mass Spectrometry/methods , Proteomics , Protein Processing, Post-Translational
15.
Immunity ; 18(2): 243-53, 2003 Feb.
Article in English | MEDLINE | ID: mdl-12594951

ABSTRACT

Upon encounter with antigen, B lymphocytes differentiate into Ig-secreting plasma cells. This step involves a massive development of secretory organelles, most notably the endoplasmic reticulum. To analyze the relationship between organelle reshaping and Ig secretion, we performed a dynamic proteomics study of B lymphoma cells undergoing in vitro terminal differentiation. By clustering proteins according to temporal expression patterns, it appeared that B cells anticipate their secretory role in a multistep process. Metabolic capacity and secretory machinery expand first to accommodate the mass production of IgM that follows.


Subject(s)
B-Lymphocytes/immunology , Immunoglobulin M/biosynthesis , Proteins/genetics , Animals , B-Lymphocytes/cytology , B-Lymphocytes/metabolism , Cell Differentiation , Cell Line , Endoplasmic Reticulum/immunology , Endoplasmic Reticulum/metabolism , Energy Metabolism , Gene Expression , Lipopolysaccharides/pharmacology , Mice , Oxidation-Reduction , Plasma Cells/cytology , Plasma Cells/immunology , Plasma Cells/metabolism , Proteomics
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