Abstract:
:As a potential tool for proteomics and protein characterization, in-gel cysteine- and arginine-specific cleavage is demonstrated by means of trypsin or endoproteinase Lys-C for six model proteins (lysozyme, alpha-lactalbumin, beta-lactoglobulin, ribonuclease A, albumin, and transferrin), ranging in size from 14 kDa to 79 kDa. Chemical modifications of cysteine (aminoethylation with bromoethylamine or N-(iodoethyl)-trifluoroacetamide, and subsequent guanidination) and lysine (acetylation) prior to tryptic digestion releases peptides delineated by cysteine or arginine residues. Peptide products are analyzed by MALDI-TOF-MS, ESI-MS, and ESI- and MALDI-MS/MS (with a quadrupole time-of-flight instrument). Complications induced by acrylamide alkylations of cysteines were avoided by substituting lower pH bis-tris polyacrylamide gels for tris-glycine. Sequence coverages from 35 to 86% were obtained and amino acid compositions of generated peptides could be confirmed by comprehensive y- and b-ion series. Detailed information about, in particular, cysteine rich proteins after gel electrophoresis were obtained. The chemistries for modification and cleavage specificities at cysteine residues provide an alternative means to characterize and identify proteins separated by gel electrophoresis.
journal_name
J Proteome Resjournal_title
Journal of proteome researchauthors
Thevis M,Ogorzalek Loo RR,Loo JAdoi
10.1021/pr025568gkeywords:
subject
Has Abstractpub_date
2003-03-01 00:00:00pages
163-72issue
2eissn
1535-3893issn
1535-3907journal_volume
2pub_type
杂志文章abstract::The fungal pathogen Fusarium graminearum (telomorph Gibberella zeae) is the causal agent of several destructive crop diseases. Identifying interactions among F. graminearum proteins and understanding their functions can provide insights into pathogenic mechanisms underlying F. graminearum-host interactions. F. gramine...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr900415b
更新日期:2009-10-01 00:00:00
abstract::New mass-tagging reagents for quantitative proteomics measurements have been designed using solid phase peptide synthesis technology. The solid phase mass tags have been used to accurately measure the relative amounts of cysteine-containing peptides in model peptide mixtures as well as in mixtures of tryptic digests i...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr034081k
更新日期:2004-01-01 00:00:00
abstract::We present METATRYP version 2 software that identifies shared peptides across the predicted proteomes of organisms within environmental metaproteomics studies to enable accurate taxonomic attribution of peptides during protein inference. Improvements include ingestion of complex sequence assembly data categories (meta...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/acs.jproteome.0c00385
更新日期:2020-11-06 00:00:00
abstract::Using an integrated transcriptomic and proteomic approach, we characterized the venom peptidome of the European red ant, Manica rubida. We identified 13 "myrmicitoxins" that share sequence similarities with previously identified ant venom peptides, one of them being identified as an EGF-like toxin likely resulting fro...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/acs.jproteome.0c00048
更新日期:2020-04-03 00:00:00
abstract::A selective method for the enrichment of O-GlcNAcylated peptides using a novel CLICK chemistry reagent is described. Peptides modified by O-GlcNAc were enzymatically labeled with N-azidoacetylgalactosamine. The azide was then reacted with a phospho-alkyne using CLICK chemistry and O-GlcNAcGalNAzPO(4)-containing peptid...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr100565j
更新日期:2011-04-01 00:00:00
abstract::Protection against injurious external insults and loss of vital fluids is essential for life and is in all organisms, from bacteria to plants and humans, provided by some form of barrier. Members of the small proline-rich (SPRR) protein family are major components of the cornified cell envelope (CE), a structure respo...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr300310b
更新日期:2012-06-01 00:00:00
abstract::A proper sample preparation, in particular, abundant protein removal is crucial in the characterization of low-abundance proteins including those harboring post-translational modifications. In human cerebrospinal fluid (CSF), approximately 80% of proteins originate from serum, and removal of major proteins is necessar...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr049750o
更新日期:2005-05-01 00:00:00
abstract::Ubiquitin ligases (E3s) serve as key regulators for the ubiquitylation-mediated pathway. The identification of the corresponding relationship between E3 and its substrates is challenging but required for understanding the regulatory network of ubiquitylation. The low abundance of ubiquitinated conjugates and high redu...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/acs.jproteome.9b00673
更新日期:2020-01-03 00:00:00
abstract::The Halobacterium salinarum gas vesicle (GV) is an extremely stable intracellular organelle with air trapped inside a proteinaceous membrane. Reported here is a comparative proteomics analysis of GV and GV depleted lysate (GVD) to reveal the membrane structural proteins. Ten proteins encoded by gvp-1 (gvpMLKJIHGFED-1 ...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr1009383
更新日期:2011-03-04 00:00:00
abstract::We have expanded our recent on-line LC-MS platform for large peptide analysis to combine collision-induced dissociation (CID), electron-transfer dissociation (ETD), and CID of an isolated charge-reduced (CRCID) species derived from ETD to determine sites of phosphorylation and glycosylation modifications, as well as t...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr070313u
更新日期:2007-11-01 00:00:00
abstract::The development of medical interventions for the preservation of disease-free longevity would be facilitated by markers that predict healthy aging. Altered protein N-glycosylation patterns have been found with increasing age and several disease states. Here we investigate whether glycans derived from the total glycopr...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr1009959
更新日期:2011-04-01 00:00:00
abstract::In-source fragmentation occurs as a byproduct of electrospray ionization. We find that ions produced as a result of in-source fragmentation often match fragment ions produced during MS/MS fragmentation, and we take advantage of this phenomenon in a novel algorithm to analyze LC-MS metabolomics data sets. Our approach ...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/acs.jproteome.8b00601
更新日期:2019-02-01 00:00:00
abstract::Francisella tularensis is a Gram-negative bacterium that causes the zoonotic disease tularemia. The historical development of tularemia as a biological weapon has led to it being characterized by the CDC as a category A biothreat agent. Neither posttranslational modification (PTM) of proteins, in particular lysine ace...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/acs.jproteome.9b00512
更新日期:2020-04-03 00:00:00
abstract::Aspergillus fumigatus is a ubiquitously distributed filamentous fungus that has emerged as one of the most serious life-threatening pathogens in immunocompromised patients. The mechanisms for its pathogenicity are poorly understood. Here, we analyzed the proteome of dormant A. fumigatus conidia as the fungal entity ha...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr9010684
更新日期:2010-07-02 00:00:00
abstract::Although the "missing protein" is a temporary concept in C-HPP, the biological information for their "missing" could be an important clue in evolutionary studies. Here we classified missing-protein-encoding genes into two groups, the genes encoding PE2 proteins (with transcript evidence) and the genes encoding PE3/4 p...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/acs.jproteome.5b00450
更新日期:2015-12-04 00:00:00
abstract::An individually darkened leaf model was used to study protein changes in the Arabidopsis mutant stay-green1 (sgr1) to partially mimic the process of leaf covering senescence that occurs naturally in the shaded rosettes of Arabidopsis plants. Utilizing this controlled and predictable induced senescence model has allowe...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr300691k
更新日期:2012-11-02 00:00:00
abstract::Target-decoy approach (TDA) is the dominant strategy for false discovery rate (FDR) estimation in mass-spectrometry-based proteomics. One of its main applications is direct FDR estimation based on counting of decoy matches above a certain score threshold. The corresponding equations are widely employed for filtering o...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/acs.jproteome.6b00144
更新日期:2017-02-03 00:00:00
abstract::Monoassociation of germfree Interleukin 10 gene deficient (IL-10-/-) 129SvEv but not wild-type mice with Enterococcus faecalis induces severe chronic colitis. Bacterial strain-specific effects on the development of chronic intestinal inflammation are not understood. We investigated the molecular mechanisms of E. faeca...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr800785m
更新日期:2009-03-01 00:00:00
abstract::Chinese hamster ovary (CHO) cell lines are widely used for the expression of therapeutic recombinant proteins, including monoclonal antibodies and other biologics. For manufacturing, cells derived from a single-cell clone are typically used to ensure product consistency. Presently, fetal bovine serum (FBS) is commonly...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr400352n
更新日期:2013-07-05 00:00:00
abstract::Butyrate, a fermentation product of the large bowel microflora, is potentially protective against the development of colorectal cancer. In vitro, butyrate has been shown to induce apoptosis and inhibit proliferation in numerous cancer cell lines, including colorectal cancer. Although these tumor suppressing properties...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr8009929
更新日期:2009-03-01 00:00:00
abstract::Reversible phosphorylations play a critical role in most biological pathways. Hence, in signaling studies great effort has been put into identification of a maximum number of phosphosites per experiment. Mass spectrometry (MS)-based phosphoproteomics approaches have been proven to be an ideal analytical method for map...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr200092z
更新日期:2011-08-05 00:00:00
abstract::A glycomic approach is developed to identify oligosaccharide markers for ovarian cancer by rapidly profiling globally released oligosaccharides. Glycoproteins shed by cancer cells are found in the supernatant (or conditioned media) of cultured cells. In this approach, shed glycoproteins are profiled for their oligosac...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr060010k
更新日期:2006-07-01 00:00:00
abstract::Acute kidney injury (AKI) is a frequent complication after cardiopulmonary bypass, but early detection of postoperative AKI remains challenging. Protein biomarkers predict AKI excellently in homogeneous cohorts but are less reliable in patients suffering from various comorbidities. We employed nuclear magnetic resonan...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/acs.jproteome.5b00219
更新日期:2015-07-02 00:00:00
abstract::Activation of AMP-activated protein kinase (AMPK) upon energy depletion stimulates energy production and limits energy utilization. Erythrocytes lacking AMPK are susceptible to suicidal cell death (eryptosis). A hallmark of eryptosis is cell membrane scrambling with phosphatidylserine exposure at the erythrocyte surfa...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr101004j
更新日期:2011-04-01 00:00:00
abstract::When analyzing proteins in complex samples using tandem mass spectrometry of peptides generated by proteolysis, the inference of proteins can be ambiguous, even with well-validated peptides. Unresolved questions include whether to show all possible proteins vs a minimal list, what to do when proteins are inferred ambi...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr200039p
更新日期:2011-07-01 00:00:00
abstract::The balance between comprehensively analyzing the proteome and using valuable mass spectrometry time is a genuine challenge in the field of proteomics. Multidimensional fractionation strategies have significantly increased proteome coverage, but often at the cost of increased mass analysis time, despite advances in ma...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/acs.jproteome.9b00082
更新日期:2019-05-03 00:00:00
abstract::Imatinib is the first molecular targeted therapy that has shown clinical success, but imatinib acquired resistance, although a rare event, is critical during the therapy of chronic myelogenous leukaemia (CML). With the aim of better understanding the molecular mechanisms accompanying acquisition of resistance to this ...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr0603708
更新日期:2007-01-01 00:00:00
abstract::In this paper, we describe a method for analyzing large-volume protein samples using capillary electrophoresis in conjunction with laser-induced fluorescence detection (CE-LIF). To improve the stacking and separation efficiencies of proteins, we added either 0.01% sodium dodecyl sulfate (SDS) or 0.01% poly(ethylene ox...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr050298y
更新日期:2006-02-01 00:00:00
abstract::The plant cell wall is a dynamic cellular compartment consisting of a complex matrix of components that can change dramatically in response to environmental stresses. During pathogen attack, for instance, a wide spectrum of proteins that participate in various sequential processes involved in plant defense is secreted...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr800649s
更新日期:2009-01-01 00:00:00
abstract::Phencyclidine (PCP), a noncompetitive N-methyl-D-aspartate receptor antagonist, induces psychotomimetic effects in humans and animals. Administration of PCP to rodents is used as a preclinical model for schizophrenia; however, the molecular mechanisms underlying the symptoms remain largely unknown. Acute PCP treatment...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr4010794
更新日期:2014-03-07 00:00:00