• 제목/요약/키워드: MALDI-TOF/TOF

검색결과 472건 처리시간 0.027초

Comparative Proteome Analysis of Celastrol-Treated Helicobacter pylori

  • Kim, Sa-Hyun
    • Biomedical Science Letters
    • /
    • 제23권4호
    • /
    • pp.395-401
    • /
    • 2017
  • Various preclinical and clinical trials have been conducted the efficacy of celastrol. In data presented in the current manuscript is the first trial to inhibit Helicobacter pylori with celastrol. In this study, the quantitative change of various H. pylori proteins including CagA and VacA by the anti-bacterial effect of celastrol was determined. The anti-H. pylori effects of celastrol was investigated by performing 2-dimensional electrophoresis and additional supporting experiments. After 2-dimensional electrophoresis analysis, spot intensities were analyzed and then each spot was identified using matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS) or peptide sequencing using Finnigan LCQ ion trap mass spectrometer (LC-MS/MS). The results show that celastrol has multiple effects on protein expression in H. pylori.

Protein Expression Profiling of Infected Murine Macrophage Cells (RAW 264.7) by Bacillus anthracis Spores

  • Seo Gwi-Moon;Nam Jeong-Ah;Oh Kwang-Gun;Chai Young-Gyu
    • Proceedings of the Microbiological Society of Korea Conference
    • /
    • 한국미생물학회 2003년도 International Meeting of the Microbiological Society of Korea
    • /
    • pp.77-79
    • /
    • 2003
  • Current therapeutic strategies far anthrax have had no significant impact on anthrax mortality over the last several decades. This study used a matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) discovery platform to generate protein expression profiles in search of overexpressed proteins in murine macrophage cells (RAW264.7) which infected with Bacillus anthracis spores as potentially novel molecular targets. Two differentially expressed proteins were identified in infected murine macrophage cells as Syndapin and CDC46, respectively. Syndapins are potential links between the cortical actin cytoskeleton and endocytosis. Other two proteins were identified from murine macrophage cells infected with avirulent spores as ITBG-2 (CD18) and HSPA5, respectively. These data demonstrate the feasibility of using a MALDI-TOF platform to generate protein expression profiles and identify potential molecular targets for anthrax therapeutics.

  • PDF

Occurrence of Microcystin-Containing Toxic Water Blooms in Central India

  • Agrawal Manish K.;Ghosh Shubhro K.;Bagchi Divya;Weckesser Juergen;Erhard Marcel;Bagchi Suvendra N.
    • Journal of Microbiology and Biotechnology
    • /
    • 제16권2호
    • /
    • pp.212-218
    • /
    • 2006
  • Three out of fourteen Microcystis-dominant cyanobacterial blooms in Central India were found to be toxic to mice ($LD_{50}$ ranging from 35-450 mg bloom dry mass/kg body weight). The liver architecture of the treated mice showed characteristic symptoms of hepatotoxicity relative to the untreated controls, with increased enzyme activities of serum lactate dehydrogenase (LDH), serum glutamate oxaloacetate transaminase (SGOT), alkaline phosphatase (ALP), and serum glutamate pyruvate transaminase (SGPT). RP-HPLC revealed the presence of microcystin-LR, microcystin-RR, and desmethyl microcystin-RR in the given region to maximum amounts of 390, 1,030, and $860{\mu}g/g$ bloom dry weight, respectively, corresponding to a maximum of 2.8 mg/l microcystin-LR in the lake water. Further confirmation of the microcystin variants was conducted using a MALDI-TOF MS analysis.

Characterization of Polysaccharides from Artemisia capillaris and Artemisia sylvatica (사철쑥과 그늘쑥의 다당류 연구)

  • 황은주;권학철;정칠만;문형인;김선여;지옥표;이강노
    • YAKHAK HOEJI
    • /
    • 제43권4호
    • /
    • pp.423-428
    • /
    • 1999
  • Two polysaccharides, ACP-UMP and ACP-ULF, were purified from the aerial part of Artemisia capillaris by anion-exchange chromatography, ultrafiltration, and gel filtration chromatography. The polysaccharides appeared to be homogenious from the results of HPLC. The molecular weights of ACP-UMF and ACP-ULF were estimated to be 16305.92 D and 3292.26 D, respectively, by MALDI-TOF MS. The sugar compositions were determined by GC to be arabinose 10.05%, xylose 1.67%, mannose 5.45G, galactose 39.06%, glucose 15.43% for ACP-UMF and arabinose 11.60%, xylose 11.15%, mannose 6.37% galactose 32.47%, glucose 18.35% for ACP-ULF. A polysaccharide, SP-M was determined to be 2462.52 D by MALDI-TOF MS. SP-M consisted mainly of rhamnose 36.49%, arabinose 29.00%, and glucose 19.38%. Incubation of CCl4-intoxicated hepatocytes with ACP-UMF reduced the levels of glutamic pyruvic transaminase (GPT) and cellular malondialdehyde (MDA) to 62.8% and 23.8%. ACP-ULF also reduced the levels of GPT and MDA to 46.1% and 38.1% and 26.3%, respectively.

  • PDF

Ubiquitin Fusion System for Recombinant Peptide Expression and Purification: Application to the Cytoplasmic Domain of Syndecan-4

  • Chae, Young-Kee;Lee, Ha-Yan;Lee, Weon-Tae
    • Bulletin of the Korean Chemical Society
    • /
    • 제28권9호
    • /
    • pp.1549-1552
    • /
    • 2007
  • The cytoplasmic domain of syndecan-4, a type I transmembrane heparan sulfate proteoglycan, was overexpressed as a fused form with the ubiquitin molecule in Escherichia coli, and the fusion protein was purified using immobilized metal affinity chromatography (IMAC). The cytoplasmic domain was released from its fusion partner by using yeast ubiquitin hydrolase (YUH), and subsequently purified by reverse phase chromatography. The integrity of the resulting peptide fragment was checked by MALDI-TOF and NMR spectroscopy. The yield of the peptide was 3.0-1.5 mg per liter in LB or minimal medium, respectively. The recombinant expression and purification of this domain will enable us its structural and functional studies using multidimensional NMR spectroscopy.

Analysis of Folate by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry

  • Cha, Sang-Won;Kim, Hie-Joon
    • Bulletin of the Korean Chemical Society
    • /
    • 제24권9호
    • /
    • pp.1308-1312
    • /
    • 2003
  • Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was used to observe folic acid and its derivatives such as tetrahydrofolate and 5-methyltetrahydrofolate in a vitamin tablet and in foods. Folic acid in a vitamin tablet was determined using angiotensin I as an internal reference. Tetrahydrofolic acid, 5-methyltetrahydrofolic acid, and an oxygenated folate were observed from a human blood sample using graphite plate. The results show that these mass spectrometric methods are useful for quickly obtaining a profile of folates.

Purification and Characterization of Moran 20K from Morus alba

  • Kim, Eun-Sun;Park, Sung-Jean;Lee, Eun-Ju;Kim, Bak-Kwang;Huh, Hoon;Lee, Bong-Jin
    • Archives of Pharmacal Research
    • /
    • 제22권1호
    • /
    • pp.9-12
    • /
    • 1999
  • A new glycoprotein was purified from the aqueous methanolic extract of the root bark of Morus alba which has been used as a component of antidiabetic remedy in Oriental Medicine. SDS-PAGE result shows that the molecular weight of the glycoprotein was approximately 20 kDa. This new glycoprotein was named as Moran 20K. The protein lowered blood glucose level in streptozotocin-induced hyperglycemic mice model and it also increased the glucose transport in cultured epididymis fat cells. The amino acid composition of the protein was analyzed, and the protein contained above 20% serine and cysteine such as insulin. The actual molecular weight of the protein was determined as 21.858 Da by MALDI-TOF mass spectroscopy.

  • PDF

Efficient Method for the Cleavage of Fullerene Oxides with Several Aromatic Amines under Ultrasonic Irradiation (초음파 조건에서 방향족 아민 화합물을 사용한 풀러렌 산화물 쪼개짐 반응)

  • Lee, Jong-Wook;Lee, Young-Min;Park, Byoung-Eun;Ko, Weon-Boe
    • Elastomers and Composites
    • /
    • 제42권1호
    • /
    • pp.1-8
    • /
    • 2007
  • The cleavage of fullerene oxides with several aromatic amines such as 4-nitroaniline, 3-nitroaniline, 4-isopropylaniline in the present of $FeCl_3$ occurred under ultrasonic irradiation in air at $25{\sim}43^{\circ}C$. The aminated fullerenes were confirmed by MALDI-TOF-MS and UV-vis spectra.

Proteome Analysis of Amniotic Fluid by gradient 2-D PAGI (Gradient 2-D PAGE를 이용한 양수 프로테옴 분석)

  • 이은희;김재찬;변상요
    • KSBB Journal
    • /
    • 제18권1호
    • /
    • pp.35-38
    • /
    • 2003
  • Analysis of proteome in amniotic fluid was performed by 2-D PAGE (polyacrylamide gel electrophoresis). Proteins in amniotic fluid were separated by centrifugation and solubilized in buffer solution for IEF, using an IPG strip of pH 4-7L. Both a homogeneous slab gel of 12.5% and a gradient gel of 8-18%, were used. After 2-D PAGE, spots were stained with silver nitrate and picked up for in-gel digestion. Digested peptides were analyzed by MALDI-TOF and proteins were further identifical. More protein spots were detected in the gradient gels and a protein not previously reported was identified.

Interaction Proteome Analysis of Xanthomonas Hrp Proteins

  • Jang, Mi;Park, Byoung-Chul;Lee, Do-Hee;Bae, Kwang-Hee;Cho, Sa-Yeon;Park, Hyun-Seok;Lee, Baek-Rak;Park, Sung-Goo
    • Journal of Microbiology and Biotechnology
    • /
    • 제17권2호
    • /
    • pp.359-363
    • /
    • 2007
  • Because of the importance of the type III protein-secretion system in bacteria-plant interaction, its function in bacterial pathogenesis of plants has been intensively studied. To identity bacterial proteins interacting with Xanthomonas hrp gene products that are involved in pathogenicity, we performed the glutathione-bead binding analysis of Xanthomonas lysates containing GST-tagged Hrp proteins. Analysis of glutathione-bead bound proteins by 1-DE and MALDI-TOF has demonstrated that Avr proteins, RecA, and several components of the type III secretion system interact with HrpB protein. This proteomic approach could provide a powerful tool in finding interaction partners of Hrp proteins whose roles in host-pathogen interaction need further studies.