• Title/Summary/Keyword: MALDI-TOF/TOF MS

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Comparative Proteomic Analysis of Virulent Korean Mycobacterium tuberculosis K-strain with Other Mycobacteria Strain Following Infection of U-937 Macrophage

  • Ryoo, Sung-Weon;Park, Young-Kil;Park, Sue-Nie;Shim, Young-Soo;Liew, Hyun-Jeong;Kang, Seong-Man;Bai, Gill-Han
    • Journal of Microbiology
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    • v.45 no.3
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    • pp.268-271
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    • 2007
  • In Korea, the Mycobacterium tuberculosis K-strain is the most prevalent clinical isolates and belongs to the Beijing family. In this study, we conducted comparative porteomics of expressed proteins of clinical isolates of the K-strain with H37Rv, H37Ra as well as the vaccine strain of Mycobacterium bovis BCG following phagocytosis by the human monocytic cell line U-937. Proteins were analyzed by 2-D PAGE and MALDI-TOF-MS. Two proteins, Mb1363 (probable glycogen phosphorylase GlgP) and MT2656 (Haloalkane dehalogenase LinB) were most abundant after phagocytosis of M. tuberculosis K-strain. This approach provides a method to determine specific proteins that may have critical roles in tuberculosis pathogenesis.

Sonochemical Reaction of Fullerene Oxides, [C70(O)n](n≥1) with Aromatic Amines (방향족 아민 화합물과 풀러렌 산화물의 [C70(O)n](n≥1)의 초음파 화학 반응)

  • Ko, Weon-Bae;Park, Byoung-Eun;Lee, Young-Min
    • Elastomers and Composites
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    • v.43 no.1
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    • pp.31-38
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    • 2008
  • Sonochemical reaction of fullerene oxides, $[C_{70}(O)_n](n\geq1)$ with several aromatic amines such as 4-nitroaniline, 3-nitroaniline, and 4-isopropylaniline, in the presence of $FeCl_3$ were investigated under ultrasonic irradiation. This method is applicable to a wide variety of aromatic amines especially ring deactivated, to afford the corresponding cleavage products under mild conditions. The aminated fullerenes were confirmed by MALDI-TOF-MS and UV-vis spectra.

Evaluation of Microbiological Contamination of Water Purifiers at Two Universities in Chungcheong Region

  • Jin Young Yun
    • Biomedical Science Letters
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    • v.29 no.4
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    • pp.256-262
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    • 2023
  • The purpose of this study is to investigate microbial contamination in water purifiers from two universities (A and B) in Chungcheong region and to evaluate about the harmfulness of the isolated bacteria to the human. The degree of microbiological contamination of six water purifiers at university A was investigated three times from July 2018 to September 2019, and nine water purifiers at university B were investigated in 2023. The isolated bacteria were biochemically identified using an API kit and Vitek-2 system, and then the bacteria were identified to the species level using MALDI-TOF MS. In addition, the possibility of human infection of the isolated bacteria was evaluated through a literature search. In July 2018 and September 2019, the number of bacteria isolated inside the faucet was below the acceptable standard for hot water, but exceed for cold water in all water purifiers. In January and September 2019, bacteria exceeding the acceptable standards were isolated nine times from the cold water of six water purifies (a total of 12 water purifiers). Bacteria identified by MALDI-TOF MS included anaerobic bacteria (Clostridium novyi, Clostridium themopalmarium etc.), Gram-positive bacilli (Microbacterium testaceum, Arthrobacter woluwensis etc.), and Gramnegative bacilli (Acinetobacter nosocomialis, Comamonas kerstersii etc.), which are difficult identify by biochemical methods. In conclusion, bacteria exceeding the acceptable standard were isolated from the cold water of most of the water purifiers. Most of the isolated bacteria were low-pathogenic bacteria from natural environment, but opportunistic bacteria that can cause infection in humans were also isolated from some water purifiers.

Synthesis of Dendrimer Based Polymeric and Macrocyclic Complexes with a Platinum-Acetylide ${\pi}-Conjugated$ Organometallic Core

  • Jang, Woo-Dong
    • Macromolecular Research
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    • v.13 no.4
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    • pp.334-338
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    • 2005
  • A three-layered poly(benzyl ether) dendrimer having a bis-ethynylbenzene core was synthesized and characterized with $^{1}H$ NMR and MALDI-TOF-MS spectroscopy. The dendrimer was reacted with platinum complexes to obtain platinum-acetylide based organometallic polymers. When the dendrimer was reacted with trans-[$PtCl_{2}(PEt_{3})_{2}$], a high molecular weight polymeric compound was formed, whereas, with cis-[$PtCl_{2}dppp$], a uniform molecular weight compound was formed, which was found to be a dimeric metallacycle by $^{1}H\;NMR,\;^{31}P\;NMR$ and ESI-TOF-MS spectroscopy. Both these complexes exhibited relatively a strong emission around 440 nm, indicating that they could be potential candidates for blue emitting polymer LEDs.

Endophytic Bacillus sp. CY22 from a Balloon Flower (Platycodon grandiflorum) Produces Surfactin Isoforms

  • Cho, Soo-Jeong;Hong, Su-Young;Kim, Jin-Young;Park, Sang-Ryeol;Kim, Min-Keun;Lim, Woo-Jin;Shin, Eun-Chule;Kim, Eun-Ju;Cho, Yong-Un;Yun, Han-Dae
    • Journal of Microbiology and Biotechnology
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    • v.13 no.6
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    • pp.859-865
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    • 2003
  • Surfactin is a mixture of cyclic lipopeptides built from variants of a heptapeptide and a ${\beta}-hydroxy$ fatty acid produced by several strains of Bacillus sp. Surfactin isoforms produced by endophytic Bacillus sp. CY22 from a balloon flower were isolated and characterized. It was found that the purified surfactin had three isoforms with protonated masses of m/z 1,008, 1,022, and 1,036, and different structures in combination with Na, K, Ca ions using MALDI-TOF MS, ESI-MS/MS, and ICP MS, respectively. In the MS/MS analysis, the isolated surfactin had the identical amino acid sequence (LLVDLL) and hydroxy fatty acids (with 13 to 15 carbons in length), even though isolated from different Bacillus strains. The sfp22 gene, required for producing the surfactin, consisted of an open reading frame (ORF) of 675 bp encoding 224 amino acid residues with a signal peptide of 20 amino acids. The predicted amino acid sequence of sfp22 was very similar to that of Ipa-8.

Biochemical mechanisms of fumigant toxicity by ethyl formate towards Myzus persicae nymphs (복숭아혹진딧물(Myzus persicae) 약충에 대한 에틸포메이트 훈증 독성의 생화학적 메커니즘)

  • Kim, Kyeongnam;Lee, Byung-Ho;Park, Jeong Sun;Yang, Jeong Oh;Lee, Sung-Eun
    • Journal of Applied Biological Chemistry
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    • v.60 no.3
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    • pp.271-277
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    • 2017
  • Ethyl formate has been used for the control of insect pests by fumigation. However, there were not many reports to show its target site of fumigant toxicity on insect pests since its first use in the agricultural industry. In the present study, we showed the presumable target sites of ethyl formate fumigation in insect pests using Myzus persicae nymphs. After ethyl formate fumigation, the nymphs of this species were collected and the changes at the biochemical and molecular level were determined. The activity of cytochrome c oxidase (COX) was approximately two-fold higher after ethyl formate fumigation. In addition, the expression levels of acetylcholinesterase (AChE) decreased gradually with increasing ethyl formate concentration. These two findings suggested that COX and AChE might be the major target sites of ethyl formate fumigation. In addition to these results, the analysis of lipid content using MALDI-TOF MS/MS identified 9 phospholipids differently generated 2-fold higher in the ethyl formate-treated nymphs than that in the control nymphs, thereby leading to changes in cell membrane composition in M. persicae nymphs. Therefore, the ethyl formate fumigation caused lethal effects on M. persicae nymphs by changing COX activity, AChE gene expression, and phospholipid production.

Proteomic Analysis of Global Changes in Protein Expression During Exposure of Gamma Radiation in Bacillus sp. HKG 112 Isolated from Saline Soil

  • Gupta, Anil Kumar;Pathak, Rajiv;Singh, Bharat;Gautam, Hemlata;Kumar, Ram;Kumar, Raj;Arora, Rajesh;Gautam, Hemant K.
    • Journal of Microbiology and Biotechnology
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    • v.21 no.6
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    • pp.574-581
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    • 2011
  • A Gram-positive bacterium was isolated from the saline soils of Jangpura (U.P.), India, and showed high-level of radiation-resistant property and survived upto 12.5 kGy dose of gamma radiation. The 16S rDNA sequence of this strain was examined, identified as Bacillus sp. strain HKG 112, and was submitted to the NCBI GenBank (Accession No. GQ925432). The mechanism of radiation resistance and gene level expression were examined by proteomic analysis of whole-cell extract. Two proteins, 38 kDa and 86.5 kDa excised from SDS-PAGE, which showed more significant changes after radiation exposure, were identified by MALDI-TOF as being flagellin and S-layer protein, respectively. Twenty selected 2-DE protein spots from the crude extracts of Bacillus sp. HKG 112, excised from 2- DE, were identified by liquid chromatography mass spectrometry (LC-MS) out of which 16 spots showed significant changes after radiation exposure and might be responsible for the radiation resistance property. Our results suggest that the different responses of some genes under radiation for the expression of radiation-dependent proteins could contribute to a physiological advantage and would be a significant initial step towards a fullsystem understanding of the radiation stress protection mechanisms of bacteria in different environments.

Proteome Analysis of Disease Resistance against Ralstonia solanacearum in Potato Cultivar CT206-10

  • Park, Sangryeol;Gupta, Ravi;Krishna, R.;Kim, Sun Tae;Lee, Dong Yeol;Hwang, Duk-ju;Bae, Shin-Chul;Ahn, Il-Pyung
    • The Plant Pathology Journal
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    • v.32 no.1
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    • pp.25-32
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    • 2016
  • Potato is one of the most important crops worldwide. Its commercial cultivars are highly susceptible to many fungal and bacterial diseases. Among these, bacterial wilt caused by Ralstonia solanacearum causes significant yield loss. In the present study, integrated proteomics and genomics approaches were used in order to identify bacterial wilt resistant genes from Rs resistance potato cultivar CT-206-10. 2-DE and MALDI-TOF/TOF-MS analysis identified eight differentially abundant proteins including glycine-rich RNA binding protein (GRP), tomato stress induced-1 (TSI-1) protein, pathogenesis-related (STH-2) protein and pentatricopeptide repeat containing (PPR) protein in response to Rs infection. Further, semi-quantitative RT-PCR identified up-regulation in transcript levels of all these genes upon Rs infection. Taken together, our results showed the involvement of the identified proteins in the Rs stress tolerance in potato. In the future, it would be interesting to raise the transgenic plants to further validate their involvement in resistance against Rs in potato.

Differential protein expression in avian liver in response to invasion by Salmonella gallinarum

  • Lee, Gang-Deog;Cho, In-Hee;So, Hyun-Kyung;Koo, Yong-bum;Lee, Jun-heon;Choi, Kang-Duk
    • Proceedings of the Korea Society of Poultry Science Conference
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    • 2004.11a
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    • pp.37-38
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    • 2004
  • Salmonella gallinarum is a pathogen that is capable of causing disease in Korean native chicken. Although Salmonella gallinarum is important world-wide pathogens of poultry, little is understood of the mechanisms of pathogenesis of Salmonella gallinarum in the chicken. This study was to investigate chicken liver proteins affected by infection of Salmonella gallinarum in Korean native chicken. The differentially expressed proteins of chicken livers were identified by using 2-dimensional electro- phoresis (2D-E) and mass spectrometry (MS). We detected more than 300 protein spots on silver stained 2D gels using pH 3∼10 gradients. Three differentially expressed protein spots were analyzed by MALDI-TOF MS and MS/MS. The obtained MS and MS/MS data were searched against a protein database using the Mascot search engine. Further researches on the identified proteins can give valuable information of mechanism of pathogenesis in chicken.

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Comparative Serum Proteomic Analysis of Serum Diagnosis Proteins of Colorectal Cancer Based on Magnetic Bead Separation and MALDI-TOF Mass Spectrometry

  • Deng, Bao-Guo;Yao, Jin-Hua;Liu, Qing-Yin;Feng, Xian-Jun;Liu, Dong;Zhao, Li;Tu, Bin;Yang, Fan
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.10
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    • pp.6069-6075
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    • 2013
  • Background: At present, the diagnosis of colorectal cancer (CRC) requires a colorectal biopsy which is an invasive procedure. We undertook this pilot study to develop an alternative method and potential new biomarkers for diagnosis, and validated a set of well-integrated tools called ClinProt to investigate the serum peptidome in CRC patients. Methods: Fasting blood samples from 67 patients diagnosed with CRC by histological diagnosis, 55 patients diagnosed with colorectal adenoma by biopsy, and 65 healthy volunteers were collected. Division was into a model construction group and an external validation group randomly. The present work focused on serum proteomic analysis of model construction group by ClinProt Kit combined with mass spectrometry. This approach allowed construction of a peptide pattern able to differentiate the studied populations. An external validation group was used to verify the diagnostic capability of the peptidome pattern blindly. An immunoassay method was used to determine serum CEA of CRC and controls. Results: The results showed 59 differential peptide peaks in CRC, colorectal adenoma and health volunteers. A genetic algorithm was used to set up the classification models. Four of the identified peaks at m/z 797, 810, 4078 and 5343 were used to construct peptidome patterns, achieving an accuracy of 100% (> CEA, P<0.05). Furthermore, the peptidome patterns could differentiate the validation group with high accuracy close to 100%. Conclusions: Our results showed that proteomic analysis of serum with MALDI-TOF MS is a fast and reproducible approach, which may provide a novel approach to screening for CRC.