Determination of the site-specific and isoform-specific glycosylation in human plasma-derived antithrombin by IEF and capillary HPLC-ESI-MS/MS.

Abstract:

:The glycan structures of the major and more than ten minor populated isoforms of antithrombin (AT) were determined after separation of the isoforms by IEF using IPG strips. The bands excised from the gel were reduced, derivatized by iodoacetamide and submitted to tryptic digestion. The digest was analyzed by RP-HPLC-ESI-MS equipped with a quadrupole ion-trap mass analyzer. MS/MS experiments allowed establishing the monosaccharide compositions in the glycopeptides. For the major isoform of alpha-AT four identical biantennary glycans with two terminal sialic acids (SA) each, a total of eight SA, were found in full agreement with the literature. In the IEF-band containing this major isoform (pI 5.18) a further, much less abundant, isoform was detected showing a fucosylation on the glycan attached to Asn155 but being of otherwise identical structure as described above. The isoforms with pI 5.10 were found to include one triantennary glycan, all antennas carrying terminal SA. The occurrence of triantennary structure is site specific, involving the peptides with Asn(135) and Asn(155), alternately. At pI 5.24 we found those four isoforms that carry the glycans like the main-isoform of alpha-AT but missing one terminal SA. There was no site specificity found for the mono-sialo structure. The isoform at pI 5.31 is the major isoform of beta-AT containing three identical biantennary structures being fully sialylated. No isoforms (above 0.5% abundance) with two glycans only or three glycans other than beta-AT were detected. Fucosylation was found in the main isoform with an abundance of about 5%, and as expected with all the other isoforms with a comparable abundance.

journal_name

Proteomics

journal_title

Proteomics

authors

Plematl A,Demelbauer UM,Josic D,Rizzi A

doi

10.1002/pmic.200401238

subject

Has Abstract

pub_date

2005-10-01 00:00:00

pages

4025-33

issue

15

eissn

1615-9853

issn

1615-9861

journal_volume

5

pub_type

杂志文章
  • Range of protein detection by selected/multiple reaction monitoring mass spectrometry in an unfractionated human cell culture lysate.

    abstract::Selected or multiple reaction monitoring is a targeted mass spectrometry method (S/MRM-MS), in which many peptides are simultaneously and consistently analyzed during a single liquid chromatography-mass spectrometry (LC-S/MRM-MS) measurement. These capabilities make S/MRM-MS an attractive method to monitor a consisten...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.201100543

    authors: Ebhardt HA,Sabidó E,Hüttenhain R,Collins B,Aebersold R

    更新日期:2012-04-01 00:00:00

  • Peroxisomal proteomics, a new tool for risk assessment of peroxisome proliferating pollutants in the marine environment.

    abstract::In an attempt to improve the detection of peroxisome proliferation as a biomarker in environmental pollution assessment, we have applied a novel approach based on peroxisomal proteomics. Peroxisomal proteins from digestive glands of mussels Mytilus galloprovincialis were analyzed using 2-DE and MS. We have generated a...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200401243

    authors: Mi J,Orbea A,Syme N,Ahmed M,Cajaraville MP,Cristóbal S

    更新日期:2005-10-01 00:00:00

  • In Depth Quantitative Proteomic and Transcriptomic Characterization of Human Adipocyte Differentiation Using the SGBS Cell Line.

    abstract::Most information on molecular processes accompanying and driving adipocyte differentiation are derived from rodent models. Here, we provide a comprehensive analysis of combined transcriptomic and proteomic alterations during adipocyte differentiation in Simpson-Golabi-Behmel Syndrome (SGBS) cells. The SGBS cells are a...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.201900405

    authors: Kalkhof S,Krieg L,Büttner P,Wabitsch M,Küntzel C,Friebe D,Landgraf K,Hanschkow M,Schubert K,Kiess W,Krohn K,Blüher M,von Bergen M,Körner A

    更新日期:2020-05-08 00:00:00

  • iTRAQ-based quantitative proteomic analysis reveals new metabolic pathways of wheat seedling growth under hydrogen peroxide stress.

    abstract::As an abundant ROS, hydrogen peroxide (H2 O2 ) plays pivotal roles in plant growth and development. In this work, we conducted for the first time an iTRAQ-based quantitative proteomic analysis of wheat seedling growth under different exogenous H2 O2 treatments. The growth of seedlings and roots was significantly restr...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.201300042

    authors: Ge P,Hao P,Cao M,Guo G,Lv D,Subburaj S,Li X,Yan X,Xiao J,Ma W,Yan Y

    更新日期:2013-10-01 00:00:00

  • Citrulline enhances myofibrillar constituents expression of skeletal muscle and induces a switch in muscle energy metabolism in malnourished aged rats.

    abstract::Citrulline (Cit) actions on muscle metabolism remain unclear. Those latter were investigated using a proteomic approach on Tibialis muscles from male Sprague-Dawley rats. At 23 months of age, rats were either fed ad libitum (AL group) or subjected to dietary restriction for 12 weeks. At the end of the restriction peri...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.201200262

    authors: Faure C,Morio B,Chafey P,Le Plénier S,Noirez P,Randrianarison-Huetz V,Cynober L,Aussel C,Moinard C

    更新日期:2013-07-01 00:00:00

  • Catch me if you can: mass spectrometry-based phosphoproteomics and quantification strategies.

    abstract::Phosphorylation of proteins is one of the most prominent PTMs and for instance a key regulator of signal transduction. In order to improve our understanding of cellular phosphorylation events, considerable effort has been devoted to improving the analysis of phosphorylation by MS-based proteomics. Different enrichment...

    journal_title:Proteomics

    pub_type: 杂志文章,评审

    doi:10.1002/pmic.201000489

    authors: Eyrich B,Sickmann A,Zahedi RP

    更新日期:2011-02-01 00:00:00

  • Quantitative assessment of the structural bias in protein-protein interaction assays.

    abstract::With recent publications of several large-scale protein-protein interaction (PPI) studies, the realization of the full yeast interaction network is getting closer. Here, we have analysed several yeast protein interaction datasets to understand their strengths and weaknesses. In particular, we investigate the effect of...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200800150

    authors: Björklund AK,Light S,Hedin L,Elofsson A

    更新日期:2008-11-01 00:00:00

  • Study of rat hypothalamic proteome by HPLC/ESI ion trap and HPLC/ESI-Q-TOF MS.

    abstract::The proteomic profile of hypothalamus, a key organ of CNS, is explored here by employing two widely used MS techniques, i.e. HPLC/ESI-ion trap and HPLC/ESI-quadrupole-TOF MS. Strong cation exchange is used for the fractionation of peptides and protein search engine MASCOT is employed for data query. One hundred and th...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.201300073

    authors: Iqbal J,Li W,Ullah K,Hasan M,Linna G,Awan U,Zhang Y,Batool S,Qing H,Deng Y

    更新日期:2013-08-01 00:00:00

  • Comparative proteomics of bacterial pathogens.

    abstract::The monitoring of gene expression via the technologies encompassed under the term 'proteomics' allows proteins of significance to be related to phenotypes associated with strain variability, environmental influences and the effects of genetic manipulation. The characterizations of these molecules are routinely perform...

    journal_title:Proteomics

    pub_type: 杂志文章,评审

    doi:10.1002/1615-9861(200104)1:4<461::AID-PROT461>3.0.

    authors: Cordwell SJ,Nouwens AS,Walsh BJ

    更新日期:2001-04-01 00:00:00

  • A glycomic approach to the identification and characterization of glycoprotein function in cells transfected with glycosyltransferase genes.

    abstract::The transfection of glycoprotein glycosyltransferase genes into cells leads to modification of both the structure and function of the glycoproteins and as a result, changes in glycome patterns. N-glycan branching enzymes hold some promise as a model system for the identification of glycome patterns. Both N-acetylgluco...

    journal_title:Proteomics

    pub_type: 杂志文章,评审

    doi:10.1002/1615-9861(200102)1:2<239::AID-PROT239>3.0.

    authors: Taniguchi N,Ekuni A,Ko JH,Miyoshi E,Ikeda Y,Ihara Y,Nishikawa A,Honke K,Takahashi M

    更新日期:2001-02-01 00:00:00

  • A DIGE approach for the assessment of rat soleus muscle changes during unloading: effect of acetyl-L-carnitine supplementation.

    abstract::After hind limb suspension, a remodeling of postural muscle phenotype is observed. This remodeling results in a shift of muscle profile from slow-oxidative to fast-glycolytic. These metabolic changes and fiber type shift increase muscle fatigability. Acetyl-L-carnitine (ALCAR) influences the skeletal muscle phenotype ...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200701176

    authors: Moriggi M,Cassano P,Vasso M,Capitanio D,Fania C,Musicco C,Pesce V,Gadaleta MN,Gelfi C

    更新日期:2008-09-01 00:00:00

  • Gel-free analysis of the human brain proteome: application of liquid chromatography and mass spectrometry on biopsy and autopsy samples.

    abstract::This paper reports on the findings of the Biomedical Research Institute, as one of the participants in the pilot study of the HUPO Brain Proteome Project. A biopsy and autopsy study sample derived from human brain was distributed among the participants for proteomic analysis. In our laboratory, attention was focused o...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200600080

    authors: Dumont D,Noben JP,Verhaert P,Stinissen P,Robben J

    更新日期:2006-09-01 00:00:00

  • Proteome analysis of proliferative response of bystander cells adjacent to cells exposed to ionizing radiation.

    abstract::Recently (Cytometry 2003, 56A, 71-80), we reported that direct cell-to-cell contact is required for stimulating proliferation of bystander rat liver cells (WB-F344) cocultured with irradiated cells, and neither functional gap junction intercellular communication nor long-range extracellular factors appear to be involv...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200600948

    authors: Gerashchenko BI,Yamagata A,Oofusa K,Yoshizato K,de Toledo SM,Howell RW

    更新日期:2007-06-01 00:00:00

  • A noncoding plant pathogen provokes both transcriptional and posttranscriptional alterations in tomato.

    abstract::Viroids are single-stranded, circular, noncoding RNAs that infect plants, causing devastating diseases. In this work, we employed 2D DIGE, followed by MS identification, to analyze the response of tomato plants infected by Citrus exocortis viroid (CEVd). Among the differentially expressed proteins detected, 45 were su...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.201200286

    authors: Lisón P,Tárraga S,López-Gresa P,Saurí A,Torres C,Campos L,Bellés JM,Conejero V,Rodrigo I

    更新日期:2013-03-01 00:00:00

  • Screening the human serum proteome for genotype-phenotype associations: an analysis of the IL6 -174G>C polymorphism.

    abstract::Interleukin (IL)-6 is a circulatory, pleiotropic cytokine with multiple roles in the immune system. Both IL-6 and the IL6 -174G>C promoter polymorphism have been linked to various diseases associated with inflammation. However, the mechanism by which the polymorphism influences disease risk is unclear. We postulated t...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200600366

    authors: Hegedus CM,Skibola CF,Bracci P,Holly EA,Smith MT

    更新日期:2007-02-01 00:00:00

  • New changes in the plasma-membrane-associated proteome of rice roots under salt stress.

    abstract::To gain a better understanding of salt stress responses in plants, we used a proteomic approach to investigate changes in rice (Oryza sativa) root plasma-membrane-associated proteins following treatment with 150 mmol/L NaCl. With or without a 48 h salt treatment, plasma membrane fractions from root tip cells of a salt...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200800340

    authors: Cheng Y,Qi Y,Zhu Q,Chen X,Wang N,Zhao X,Chen H,Cui X,Xu L,Zhang W

    更新日期:2009-06-01 00:00:00

  • In macropore tryptic digestion at acidic pH and its implication for proteomics.

    abstract::Gan et al. (Proteomics 2013, 13, 3117-3123) described a new "macropore" protocol for effective protein digestion by trypsin suitable for a wide range of pH including acidic pH. It was effective not only in experiments with solutions of a model protein (myoglobin), but also with a subfraction of rat liver cytosol. This...

    journal_title:Proteomics

    pub_type: 评论,杂志文章

    doi:10.1002/pmic.201300440

    authors: Medvedev AE

    更新日期:2013-11-01 00:00:00

  • Fluorophore-assisted light inactivation: a high-throughput tool for direct target validation of proteins.

    abstract::To exploit advances in proteomics for drug discovery, high-throughout methods for target validation that directly address the cellular roles of proteins are required. To do this, we have characterized fluorophore-assisted light inactivation (FALI) which uses coherent or diffuse light targeted by fluorescein-labeled pr...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/1615-9861(200203)2:3<247::aid-prot247>3.0.

    authors: Beck S,Sakurai T,Eustace BK,Beste G,Schier R,Rudert F,Jay DG

    更新日期:2002-03-01 00:00:00

  • Profiling MS proteomics data using smoothed non-linear energy operator and Bayesian additive regression trees.

    abstract::This paper proposes a novel profiling method for SELDI-TOF and MALDI-TOF MS data that integrates a novel peak detection method based on modified smoothed non-linear energy operator, correlation-based peak selection and Bayesian additive regression trees. The peak detection and classification performance of the propose...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200800502

    authors: He S,Li X,Viant MR,Yao X

    更新日期:2009-09-01 00:00:00

  • Proteomic characterization of human serum proteins associated with the fat-derived hormone adiponectin.

    abstract::Adiponectin is a fat cell-secreted hormone with antidiabetic and anti-inflammatory activities. The reduced adiponectin levels are associated with obesity-related metabolic syndrome. Replenishment of this hormone into animal models can improve insulin sensitivity, decrease blood glucose and lipid levels, and prevent th...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200500840

    authors: Wang Y,Xu LY,Lam KS,Lu G,Cooper GJ,Xu A

    更新日期:2006-07-01 00:00:00

  • Proteomic Biomarkers to Guide Stratification for Covid-19 Treatment: Exemplifying a Path Forward Toward Implementation?

    abstract::Proteins and peptides serve as biomarkers in the context of multiple pathologies. The hypothesis that protein or peptide biomarkers may also be of value in the context of the Covid-19 pandemic appears self-evident. Proteome based biomarkers are not expected to display significant added value in the detection of viral ...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.202000229

    authors: Jankowski J

    更新日期:2020-12-01 00:00:00

  • Descriptive proteomic analysis shows protein variability between closely related clinical isolates of Mycobacterium tuberculosis.

    abstract::The use of isobaric tags such as iTRAQ allows the relative and absolute quantification of hundreds of proteins in a single experiment for up to eight different samples. More classical techniques such as 2-DE can offer a complimentary approach for the analysis of complex protein samples. In this study, the proteomes of...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200900836

    authors: Mehaffy C,Hess A,Prenni JE,Mathema B,Kreiswirth B,Dobos KM

    更新日期:2010-05-01 00:00:00

  • Comparative proteome analysis of porcine follicular fluid and serum reveals that excessive alpha(2)-macroglobulin in serum hampers successful expansion of cumulus-oocyte complexes.

    abstract::Porcine follicular fluid (pFF) constitutes the micro-environment of the maturing oocyte. Although pFF is a transudate of serum, in pigs, it is superior to serum in promoting in vitro expansion of the cumulus cells, a specialized cell population surrounding the oocyte. A comparative proteome analysis of autologous seru...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200900270

    authors: Bijttebier J,Tilleman K,Dhaenens M,Deforce D,Van Soom A,Maes D

    更新日期:2009-10-01 00:00:00

  • Torches, Candles, Lamps, Lanterns, Flashlights, Spotlights, Night Vision Goggles… You Need Them All to See in Darkness.

    abstract::Articles assembled in the second part of this Special Issue describe some experimental and computational approaches for the structural and functional characterization of intrinsically disordered proteins. Since these tools represent specialized gear for the focused analysis of various aspects of dark proteome, they ca...

    journal_title:Proteomics

    pub_type: 社论

    doi:10.1002/pmic.201900085

    authors: Uversky VN

    更新日期:2019-03-01 00:00:00

  • Membrane-Based SDS Depletion Ahead of Peptide and Protein Analysis by Mass Spectrometry.

    abstract::SDS interferes with both bottom-up and top-down MS analysis, requiring removal prior to detection. Filter-aided sample preparation (FASP) is favored for bottom-up proteomics (BUP) while acetone precipitation is popular for top-down proteomics (TDP). We recently demonstrated acetone precipitation in a membrane filter c...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.201700025

    authors: Unterlander N,Doucette AA

    更新日期:2018-05-01 00:00:00

  • Consequences of the discontinuation of the International Protein Index (IPI) database and its substitution by the UniProtKB "complete proteome" sets.

    abstract::The International Protein Index (IPI) database has been one of the most widely used protein databases in MS proteomics approaches. Recently, the closure of IPI in September 2011 was announced. Its recommended replacement is the new UniProt Knowledgebase (UniProtKB) "complete proteome" sets, launched in May 2011. Here,...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.201100363

    authors: Griss J,Martín M,O'Donovan C,Apweiler R,Hermjakob H,Vizcaíno JA

    更新日期:2011-11-01 00:00:00

  • An interaction proteomics survey of transcription factor binding at recurrent TERT promoter mutations.

    abstract::Aberrant telomerase reactivation in differentiated cells represents a major event in oncogenic transformation. Recurrent somatic mutations in the human telomerase reverse transcriptase (TERT) promoter region, predominantly localized to two nucleotide positions, are highly prevalent in many cancer types. Both mutations...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.201500327

    authors: Makowski MM,Willems E,Fang J,Choi J,Zhang T,Jansen PW,Brown KM,Vermeulen M

    更新日期:2016-02-01 00:00:00

  • Redox Proteomic Analysis Reveals Oxidative Modifications of Proteins by Increased Levels of Intracellular Reactive Oxygen Species during Hypoxia Adaptation of Aspergillus fumigatus.

    abstract::Aspergillus fumigatus faces abrupt changes in oxygen concentrations at the site of infection. An increasing number of studies has demonstrated that elevated production of intracellular reactive oxygen species (ROS) under low oxygen conditions plays a regulatory role in modulating cellular responses for adaptation to h...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.201800339

    authors: Shekhova E,Ivanova L,Krüger T,Stroe MC,Macheleidt J,Kniemeyer O,Brakhage AA

    更新日期:2019-03-01 00:00:00

  • An improved method of sample preparation on AnchorChip targets for MALDI-MS and MS/MS and its application in the liver proteome project.

    abstract::An improved method for sample preparation for MALDI-MS and MS/MS using AnchorChip targets is presented. The method, termed the SMW method (sample, matrix wash), results in better sensitivity for peptide mass fingerprinting as well as for sequencing by MS/MS than previously published methods. The method allows up-conce...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200600184

    authors: Zhang X,Shi L,Shu S,Wang Y,Zhao K,Xu N,Liu S,Roepstorff P

    更新日期:2007-07-01 00:00:00

  • Comprehensive two-dimensional liquid chromatography mass spectrometric profiling of the rat hippocampal proteome.

    abstract::In this study, we performed the first high-throughput and comprehensive proteomic profiling of the rat hippocampal proteome. Using a combination of 2-D LC-MS and data analysis with the Rosetta Elucidator(®) system, we identified 1340 unique proteins. Functional classification showed that most of these were associated ...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.201000525

    authors: Yang X,Levin Y,Rahmoune H,Ma D,Schöffmann S,Umrania Y,Guest PC,Bahn S

    更新日期:2011-02-01 00:00:00