A detergent- and cyanogen bromide-free method for integral membrane proteomics: application to Halobacterium purple membranes and the human epidermal membrane proteome.

Abstract:

:A simple and rapid method for characterizing hydrophobic integral membrane proteins and its utility for membrane proteomics using microcapillary liquid chromatography coupled on-line with tandem mass spectrometry (microLC-MS/MS) is described. The present technique does not rely on the use of detergents, strong organic acids or cyanogen bromide-mediated proteolysis. A buffered solution of 60% methanol was used to extract, solubilize, and tryptically digest proteins within a preparation of Halobacterium (H.) halobium purple membranes. Analysis of the digested purple membrane proteins by microLC-MS/MS resulted in the identification of all the predicted tryptic peptides of bacteriorhodopsin, including those that are known to be post-translationally modified. In addition, 40 proteins from the purple membrane preparation were also identified, of which 80% are predicted to contain between 1 and 16 transmembrane domains. To evaluate the general applicability of the method, the same extraction, solubilization, and digestion conditions were applied to a plasma membrane fraction prepared from human epidermal sheets. A total of 117 proteins was identified in a single microLC-MS/MS analysis, of which 55% are known to be integral or associated with the plasma membrane. Due to its simplicity, efficiency, and absence of MS interfering compounds, this technique can be used for the characterization of other integral membrane proteins and may be concomitantly applied for the analysis of membrane protein complexes or large-scale proteomic studies of different membrane samples.

journal_name

Proteomics

journal_title

Proteomics

authors

Blonder J,Conrads TP,Yu LR,Terunuma A,Janini GM,Issaq HJ,Vogel JC,Veenstra TD

doi

10.1002/pmic.200300543

subject

Has Abstract

pub_date

2004-01-01 00:00:00

pages

31-45

issue

1

eissn

1615-9853

issn

1615-9861

journal_volume

4

pub_type

杂志文章
  • A peptide resource for the analysis of Staphylococcus aureus in host-pathogen interaction studies.

    abstract::Staphylococcus aureus is an opportunistic human pathogen, which can cause life-threatening disease. Proteome analyses of the bacterium can provide new insights into its pathophysiology and important facets of metabolic adaptation and, thus, aid the recognition of targets for intervention. However, the value of such pr...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.201500091

    authors: Depke M,Michalik S,Rabe A,Surmann K,Brinkmann L,Jehmlich N,Bernhardt J,Hecker M,Wollscheid B,Sun Z,Moritz RL,Völker U,Schmidt F

    更新日期:2015-11-01 00:00:00

  • Quantitative proteomics reveals differentially expressed proteins in murine preneoplastic intestine in a model of intestinal tumorigenesis induced by low dietary folate and MTHFR deficiency.

    abstract::Colorectal cancer risk is increased when dietary folate intake is low, with or without a deficiency in methylenetetrahydrofolate reductase (MTHFR). We have observed that intestinal tumors are induced in mice fed low-folate diets, and that tumor incidence is increased when these mice also have MTHFR deficiency. This st...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.201400280

    authors: Leclerc D,Dejgaard K,Mazur A,Deng L,Wu Q,Nilsson T,Rozen R

    更新日期:2014-11-01 00:00:00

  • A nascent proteome study combining click chemistry with 2DE.

    abstract::To investigate the dynamic cellular response to a condition change, selective labeling of the nascent proteome is necessary. Here, we report a method combining click chemistry protein labeling with 2D DIGE. To test the relevance of the method, we compared nascent proteomes of actively growing bacterial cells with that...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.201200393

    authors: Osterman IA,Ustinov AV,Evdokimov DV,Korshun VA,Sergiev PV,Serebryakova MV,Demina IA,Galyamina MA,Govorun VM,Dontsova OA

    更新日期:2013-01-01 00:00:00

  • Highly Efficient Identification of O-GalNAc Glycosylation by an Acid-Assisted Glycoform Simplification Approach.

    abstract::Compared with N-linked glycosylation, the analysis of O-GalNAc glycosylation is extremely challenging due to the high structure diversity of glycans and lack of glycosidases to release O-GalNAc glycans. In this work, a glycoform simplification strategy by combining HILIC enrichment with chemical de-sialylation to char...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.201800042

    authors: You X,Qin H,Mao J,Tian Y,Dong M,Guo Z,Liang X,Wang L,Jin Y,Ye M

    更新日期:2018-09-01 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

  • The New Face of the Lipid Droplet: Lipid Droplet Proteins.

    abstract::The lipid droplet (LD) is an organelle with vital functions found in nearly all organisms. LD proteomic research has provided fundamentally important insights into this organelle's functions. The review provides a summary of LD proteomic studies conducted across diverse organisms and cell and tissue types. The accumul...

    journal_title:Proteomics

    pub_type: 杂志文章,评审

    doi:10.1002/pmic.201700223

    authors: Zhang C,Liu P

    更新日期:2019-05-01 00:00:00

  • Unrestricted identification of modified proteins using MS/MS.

    abstract::Proteins undergo PTM, which modulates their structure and regulates their function. Estimates of the PTM occurrence vary but it is safe to assume that there is an important gap between what is currently known and what remains to be discovered. The highest throughput and most comprehensive efforts to catalogue protein ...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200900502

    authors: Ahrné E,Müller M,Lisacek F

    更新日期:2010-02-01 00:00:00

  • Genome-scale identification of UDP-GlcNAc-dependent pathways.

    abstract::Metabolite flux to UDP-GlcNAc and Golgi N-glycan biosynthesis regulates surface residency of glycoprotein receptors and transporters, and thus sensitivities of cells to extracellular cues. Salvage of GlcNAc increases UDP-GlcNAc and branching of N-glycans progressively, but displays an optimum for cell proliferation an...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200800208

    authors: Lau KS,Khan S,Dennis JW

    更新日期:2008-08-01 00:00:00

  • Identification of proteins involved in inhibition of spheroid formation under microgravity.

    abstract::Many types of cells transit in vitro from a two- to a three-dimensional growth, when they are exposed to microgravity. The underlying mechanisms are not yet understood. Hence, we investigated the impact of microgravity on protein content and growth behavior. For this purpose, the human thyroid cancer cells FTC-133 wer...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.201500067

    authors: Riwaldt S,Pietsch J,Sickmann A,Bauer J,Braun M,Segerer J,Schwarzwälder A,Aleshcheva G,Corydon TJ,Infanger M,Grimm D

    更新日期:2015-09-01 00:00:00

  • Native immunoblotting of blue native gels to identify conformation-specific antibodies.

    abstract::A large repertoire of immunological methods permits monitoring the interaction of antibodies with their specific antigen. However, recognition of a protein by a conformation-specific antibody represents a challenge because native conditions must be kept throughout the assay. Native immunoblotting of blue native gels c...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200900468

    authors: Zickermann V,Wumaier Z,Wrzesniewska B,Hunte C,Schägger H

    更新日期:2010-01-01 00:00:00

  • 6th HUPO Annual World Congress - Proteomics Standards Initiative Workshop 6-10 October 2007, Seoul, Korea.

    abstract::The plenary session of the Proteomics Standards Initiative of the Human Proteome Organisation at the 6th annual HUPO world congress updated the delegates on the current status of the ongoing work of this group. The new MS interchange format, mzML, had been entered into the HUPO document process immediately prior to th...

    journal_title:Proteomics

    pub_type:

    doi:10.1002/pmic.200701086

    authors: Orchard S,Martens L,Tasman J,Binz PA,Albar JP,Hermjakob H

    更新日期:2008-04-01 00:00:00

  • Assignment of protein function and discovery of novel nucleolar proteins based on automatic analysis of MEDLINE.

    abstract::Attribution of the most probable functions to proteins identified by proteomics is a significant challenge that requires extensive literature analysis. We have developed a system for automated prediction of implicit and explicit biologically meaningful functions for a proteomics study of the nucleolus. This approach u...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200600693

    authors: Schuemie M,Chichester C,Lisacek F,Coute Y,Roes PJ,Sanchez JC,Kors J,Mons B

    更新日期:2007-03-01 00:00:00

  • Dihydroxyacetone-induced death is accompanied by advanced glycation endproduct formation in selected proteins of Saccharomyces cerevisiae and Caenorhabditis elegans.

    abstract::Advanced glycation endproduct (AGE) formation is an important mechanism for protein deterioration during diabetic complications and ageing. The effects on AGE formation following dihydroxyacetone (DHA) stress were studied in two model organisms, the yeast Saccharomyces cerevisiae and the nematode Caenorhabditis elegan...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200700165

    authors: Molin M,Pilon M,Blomberg A

    更新日期:2007-10-01 00:00:00

  • Stage-dependent increase of orosomucoid and zinc-alpha2-glycoprotein in urinary bladder cancer.

    abstract::Identification and characterization of biomarkers in body fluids such as serum or urine serve as a basis for early detection of diseases, particularly of cancer. Performing 2-DE with subsequent MS analyses, conventional immunoblotting and immunohistochemistry we identified two proteins, orosomucoid (ORM) and human zin...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200402005

    authors: Irmak S,Tilki D,Heukeshoven J,Oliveira-Ferrer L,Friedrich M,Huland H,Ergün S

    更新日期:2005-11-01 00:00:00

  • Proteomic profiling of extracellular vesicles secreted from Toxoplasma gondii.

    abstract::Toxoplasma gondii infects a wide range of hosts worldwide, including humans and domesticated animals causing toxoplasmosis disease. Recently, exosomes, small extracellular vesicles (EV) that contain nucleic acids, proteins, and lipids derived from their original cells were linked with disease protection. The effect of...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.201600477

    authors: Wowk PF,Zardo ML,Miot HT,Goldenberg S,Carvalho PC,Mörking PA

    更新日期:2017-08-01 00:00:00

  • Quantitative chemical proteomics in small-scale culture of phorbol ester stimulated basal breast cancer cells.

    abstract::Basal-like breast cancers are commonly negative for expression of estrogen and progesterone receptors and HER-2 (triple-negative breast cancer), which makes this subtype of breast cancers more aggressive and less responsive to standard treatment. We have applied a small-scale chemical proteomics method using bisindoly...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.201000801

    authors: Dolai S,Xu Q,Liu F,Molloy MP

    更新日期:2011-07-01 00:00:00

  • A rare and often unrecognized cerebromeningitis and hemodynamic disorder: a major cause of sudden death in somatic cell cloned piglets.

    abstract::In this study, we generated 40 somatic cell cloned (scNT) piglets. Of these, five piglets were stillborn, 22 scNT piglets died suddenly within the first week of life, and 1 piglet died after 40 days. Twelve scNT piglets are still healthy. The birth weights of compromised scNT piglets in comparison with those of normal...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200401079

    authors: Park MR,Cho SK,Lee SY,Choi YJ,Park JY,Kwon DN,Son WJ,Paik SS,Kim T,Han YM,Kim JH

    更新日期:2005-05-01 00:00:00

  • Proteomic characterization of the major arthropod associates of the carnivorous pitcher plant Sarracenia purpurea.

    abstract::The array of biomolecules generated by a functioning ecosystem represents both a potential resource for sustainable harvest and a potential indicator of ecosystem health and function. The cupped leaves of the carnivorous pitcher plant, Sarracenia purpurea, harbor a dynamic food web of aquatic invertebrates in a fully ...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.201000256

    authors: Gotelli NJ,Smith AM,Ellison AM,Ballif BA

    更新日期:2011-06-01 00:00:00

  • Difference gel electrophoresis.

    abstract::DIGE is a protein labelling and separation technique allowing quantitative proteomics of two or more samples by optical fluorescence detection of differentially labelled proteins that are electrophoretically separated on the same gel. DIGE is an alternative to quantitation by MS-based methodologies and can circumvent ...

    journal_title:Proteomics

    pub_type: 杂志文章,评审

    doi:10.1002/pmic.200800298

    authors: Timms JF,Cramer R

    更新日期:2008-12-01 00:00:00

  • Immune proteomics of Staphylococcus aureus.

    abstract::Immune proteomics is an increasingly powerful tool for the investigation of the adaptive immune response to natural encounters between micro-organisms and their hosts. The versatile species Staphylococcus aureus serves to illustrate how these techniques can be employed to appreciate the complexity and diversity of the...

    journal_title:Proteomics

    pub_type: 杂志文章,评审

    doi:10.1002/pmic.201100010

    authors: Bröker BM,van Belkum A

    更新日期:2011-08-01 00:00:00

  • Proteomic analysis of individual variation in normal livers of human beings using difference gel electrophoresis.

    abstract::Normal Chinese Liver Proteome Expression Profile is one of the major parts of Human Liver Proteome Project. Before starting the studies, it is necessary to examine the interindividual variation of normal liver proteome and evaluate the minimal size of samples for proteomic analysis. In this study, normal liver samples...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200600006

    authors: Zhang X,Guo Y,Song Y,Sun W,Yu C,Zhao X,Wang H,Jiang H,Li Y,Qian X,Jiang Y,He F

    更新日期:2006-10-01 00:00:00

  • An improved mechanically durable electrophoresis gel matrix that is fully compatible with fluorescence-based protein detection technologies.

    abstract::Unfortunately, conventional large-format polyacrylamide gels are mechanically fragile, often tearing during the subsequent manipulations required for visualization of the proteins. This problem is compounded when large-format two-dimensional gels are subjected to multiple staining procedures in order to detect differe...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.200300440

    authors: Schulenberg B,Arnold B,Patton WF

    更新日期:2003-07-01 00:00:00

  • Proteome analysis of differentially displayed proteins as a tool for investigating ozone stress in rice (Oryza sativa L.) seedlings.

    abstract::Employing classical two-dimensional electrophoresis (2-DE), amino acid sequencing and immunoblot analysis, we examine for the first time the effect of ozone, a highly notorious environmental pollutant, on rice seedling proteins. Drastic visible necrotic damage to leaf by ozone and consequent increase in ascorbate pero...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/1615-9861(200208)2:8<947::AID-PROT947>3.0.

    authors: Agrawal GK,Rakwal R,Yonekura M,Kubo A,Saji H

    更新日期:2002-08-01 00:00:00

  • Quantitative proteomic analysis to decipher the differential apoptotic response of bortezomib-treated APL cells before and after retinoic acid differentiation reveals involvement of protein toxicity mechanisms.

    abstract::The ubiquitin-proteasome system allows the targeted degradation of proteins and plays a critical role in the regulation of many cellular processes. Proteasome inhibition is a recent antitumor therapeutic strategy and bortezomib was the first proteasome inhibitor approved for clinical use. In this study, we used the NB...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.201200233

    authors: Uttenweiler-Joseph S,Bouyssié D,Calligaris D,Lutz PG,Monsarrat B,Burlet-Schiltz O

    更新日期:2013-01-01 00:00:00

  • Hierarchical grid transformation for image warping in the analysis of two-dimensional electrophoresis gels.

    abstract::Two-dimensional electrophoresis is a widely used method for separating a large number of proteins from complex protein mixtures and for revealing differential patterns of protein expressions. In the computer-assisted proteome research, the comparison of protein separation profiles involves several heuristic steps, ran...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/1615-9861(200211)2:11<1504::AID-PROT1504>3

    authors: Salmi J,Aittokallio T,Westerholm J,Griese M,Rosengren A,Nyman TA,Lahesmaa R,Nevalainen O

    更新日期:2002-11-01 00:00:00

  • Proteomic Analysis of Endothelial Cells Exposed to Ultrasmall Nanoparticles Reveals Disruption in Paracellular and Transcellular Transport.

    abstract::The large interactive surfaces of nanoparticles (NPs) increase the opportunities to develop NPs for vascular targeting. Proteomic analysis of endothelial cells exposed to NPs reveals the cellular response and turns the focus into the impairment of the endothelial permeability. Here, quantitative proteomics and transcr...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.201800228

    authors: Kuruvilla J,Bayat N,Cristobal S

    更新日期:2019-03-01 00:00:00

  • Expanding the mouse embryonic stem cell proteome: combining three proteomic approaches.

    abstract::The current study used three different proteomic strategies, which differed by their extent of intact protein separation, to examine the proteome of a pluripotent mouse embryonic stem cell line, R1. Proteins from whole-cell lysates were subjected either to 2-D-LC, or 1-DE, or were unfractionated prior to enzymatic dig...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.201000039

    authors: Gundry RL,Tchernyshyov I,Sheng S,Tarasova Y,Raginski K,Boheler KR,Van Eyk JE

    更新日期:2010-07-01 00:00:00

  • RNAi Transfection Results in Lipidome Changes.

    abstract::RNAi experiments are ubiquitously used in cell biology and are achieved by transfection of small interfering RNAs (siRNAs) into cells using a transfection reagent. These results in knock-down of proteins of interest, and the phenotypic consequences are then analyzed. It is reported here that two common RNA interferenc...

    journal_title:Proteomics

    pub_type: 杂志文章

    doi:10.1002/pmic.201800298

    authors: Özbalci C,Storck EM,Eggert US

    更新日期:2019-07-01 00:00:00

  • Proteomics of aluminum tolerance in plants.

    abstract::Aluminum (Al) toxicity is a major constraint for plant root development and growth as well as crop yield in acidic soils, which constitute approximately 40% of the potentially arable lands worldwide. The mechanisms of Al tolerance in plants are not well understood. As a whole systems approach, proteomic techniques hav...

    journal_title:Proteomics

    pub_type: 杂志文章,评审

    doi:10.1002/pmic.201300252

    authors: Zheng L,Lan P,Shen RF,Li WF

    更新日期:2014-03-01 00:00:00

  • Has the yo-yo stopped? An assessment of human protein-coding gene number.

    abstract::Since the identification of approximately 25,000 proteins from the draft human genome assembly in 2001, estimates of the total have oscillated between 30,000 and 70,000. The recently announced genome closure has not generated a consensus gene count despite this being a key parameter for many areas of biology including...

    journal_title:Proteomics

    pub_type: 杂志文章,评审

    doi:10.1002/pmic.200300700

    authors: Southan C

    更新日期:2004-06-01 00:00:00