Profiling ofCaenorhabditis elegans proteins using two-dimensional gel electrophoresis and matrix assisted laser desorption/ionization-time of flight-mass spectrometry

Kaji, Hiroyuki; Tsuji, Takahisa; Mawuenyega, Kwasi G.; Wakamiya, Akiko; Taoka, Masato; Isobe, Toshiaki

doi:10.1002/(sici)1522-2683(20000501)21:9<1755::aid-elps1755>3.0.co;2-s

Cited by 50 publications

(11 citation statements)

References 21 publications

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“…To further identify protein spots that were otherwise undetectable by either N-terminal amino acid analysis or immunoblotting, in-gel digestion was performed by modifying the procedures described by Kaji et al [30]. Protein spots excised from the Coomassie blue stained gel were destained in microtubes with 0.2 mL acetonitrile for 15 min and dried completely in a centrifugal evaporator.…”

Section: In-gel Digestionmentioning

confidence: 99%

Proteomic analysis of lipopolysaccharide-induced apoptosis in PC12 cells

et al. 2002

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We employed rat pheochromocytoma PC12 cells as our model system to identify cellular proteins that accompany Escherichia coli lipopolysaccharide (LPS)-induced apoptosis, based on a proteomic approach. Cell viability tests revealed that naïve PC12 cells underwent cell death in a dose-dependent manner after treatment with LPS. Flow cytometric analysis confirmed that apoptosis was primarily responsible for the observed cell death. Two-dimensional electrophoresis in conjunction with N-terminal sequencing, immunoblot, matrix-assisted laser desorption/ionization-time of flight analysis or computer matching with protein databases further revealed that the LPS-induced apoptosis is accompanied by an augmented level of calreticulin, calcium binding protein 50, endoplasmic reticulum protein 60 (ERP60), heat shock protein 60 (HSP60) or HSP90, and a reduced level of amphoterin, cytochrome c oxidase polypeptide VIa-liver or ERP29. These proteins are associated with endoplasmic reticulum, mitochondria or cell membrane, and are with known or potential roles in apoptosis. Their identification therefore provides an impetus for further delineation of the cellular and molecular basis of apoptotic cell death and sepsis based on proteomic profiling of PC12 cells.

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Section: In-gel Digestionmentioning

confidence: 99%

Proteomic analysis of lipopolysaccharide-induced apoptosis in PC12 cells

et al. 2002

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“…We could also identify proteins localized in membranes like ATP synthase (Y49A3A.2) in lysosomes or vacuolar ATP synthase F20B6.2 and proteins localized in the nucleus like ZK418.9 (RNA binding protein) and C41C4.8 (P97 protein). Five of the proteins on our map (F25H2.11, F25H2.10, C18A11.7, K04D7.1, T21B10.2), were identified earlier by Edman degradation [3] and 18 proteins by MALDI-TOF [6]. Differences in theoretical pI and M r values are due to usage of different databases.…”

Section: Discussionmentioning

confidence: 89%

“…Moreover, it was the first multicellular organism with a completely sequenced genome, a valuable prerequisite to establish C. elegans as a new model organism in proteomics. We constructed a 2-D proteome [3] and 18 proteins by MALDI-TOF [6]. Differences in theoretical pI and M r values are due to usage of different databases.…”

Section: Discussionmentioning

confidence: 99%

“…The protein expression patterns of worms of different ages were compared [2] and after separating proteins from a mixed worm population, 12 spots were identified by Edman degradation [3,4]. Aspartyl proteases of C. elegans were analyzed on 2-D gels [5] and, in a recent publication, the profiling of C. elegans proteins by 2-DE and MALDI-TOF yielded the identification of 69 spots after IEF on 3-10 NL gradients [6]. 2-D protein maps of microorganisms like Saccharomyces [7][8][9], E. coli [10] and Haemophilus [11] were constructed and the protein expression of different body parts of Drosophila [12,13], the second multicellular organism with a recently completely sequenced genome, has been analyzed and anno-tated on 2-D gels.…”

Section: Introductionmentioning

confidence: 99%

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A two-dimensional protein map ofCaenorhabditis elegans

et al. 2001

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A protein map of Caenorhabditis elegans was constructed by using two-dimensional gel electrophoresis followed by peptide mass fingerprinting. A whole worm extract of a mixed population was separated on immobilized pH gradient strips 4-7 L, 3-10 NL, 6-11 L and 12% sodium dodecyl sulfate-polyacrylamide gel eletrophoresis (SDS-PAGE) gels. Gels were stained with colloidal Coomassie blue and 286 spots representing 152 proteins were subsequently identified by matrix-assisted laser desorption/ionization-mass spectrometry after in-gel digestion with trypsin. Most of the identified proteins with known cellular function were enzymes related to carbohydrate and lipid metabolism, or structural proteins with subcellular locations in the cytoplasm, mitochondria or cytoskeleton.

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“…and some narrative reviews mentioning the lack of data on MALDI-TOF in helminthology [32, 66, 67]. Yet, most studies focused on distinct analyses of specific components, such as peptides [66–86], proteins [69, 87–114], lipids [61, 62, 115–124], carbohydrates [125–143] and nucleic acids [144] in a research context. Hence, MALDI-TOF was mainly applied to study and compare the proteome or the peptidome of different helminth species, and most reports focused on C. elegans .…”

Section: Resultsmentioning

confidence: 99%

MALDI-TOF mass spectrometry as a diagnostic tool in human and veterinary helminthology: a systematic review

et al. 2019

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Background Matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry (MS) has become a widely used technique for the rapid and accurate identification of bacteria, mycobacteria and certain fungal pathogens in the clinical microbiology laboratory. Thus far, only few attempts have been made to apply the technique in clinical parasitology, particularly regarding helminth identification. Methods We systematically reviewed the scientific literature on studies pertaining to MALDI-TOF MS as a diagnostic technique for helminths (cestodes, nematodes and trematodes) of medical and veterinary importance. Readily available electronic databases (i.e. PubMed/MEDLINE, ScienceDirect, Cochrane Library, Web of Science and Google Scholar) were searched from inception to 10 October 2018, without restriction on year of publication or language. The titles and abstracts of studies were screened for eligibility by two independent reviewers. Relevant articles were read in full and included in the systematic review. Results A total of 84 peer-reviewed articles were considered for the final analysis. Most papers reported on the application of MALDI-TOF for the study of Caenorhabditis elegans , and the technique was primarily used for identification of specific proteins rather than entire pathogens. Since 2015, a small number of studies documented the successful use of MALDI-TOF MS for species-specific identification of nematodes of human and veterinary importance, such as Trichinella spp. and Dirofilaria spp. However, the quality of available data and the number of examined helminth samples was low. Conclusions Data on the use of MALDI-TOF MS for the diagnosis of helminths are scarce, but recent evidence suggests a potential role for a reliable identification of nematodes. Future research should explore the diagnostic accuracy of MALDI-TOF MS for identification of (i) adult helminths, larvae and eggs shed in faecal samples; and (ii) helminth-related proteins that are detectable in serum or body fluids of infected individuals. Electronic supplementary material The online version of this article (10.1186/s13071-019-3493-9) contains supplementary material, which is available to authorized users.

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Profiling ofCaenorhabditis elegans proteins using two-dimensional gel electrophoresis and matrix assisted laser desorption/ionization-time of flight-mass spectrometry

Cited by 50 publications

References 21 publications

Proteomic analysis of lipopolysaccharide-induced apoptosis in PC12 cells

Proteomic analysis of lipopolysaccharide-induced apoptosis in PC12 cells

A two-dimensional protein map ofCaenorhabditis elegans

MALDI-TOF mass spectrometry as a diagnostic tool in human and veterinary helminthology: a systematic review

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