• 제목/요약/키워드: MALDI-TOF MS analysis

검색결과 166건 처리시간 0.024초

Identification of Protein Markers Specific for Papillary Renal Cell Carcinoma Using Imaging Mass Spectrometry

  • Na, Chan Hyun;Hong, Ji Hye;Kim, Wan Sup;Shanta, Selina Rahman;Bang, Joo Yong;Park, Dongmin;Kim, Hark Kyun;Kim, Kwang Pyo
    • Molecules and Cells
    • /
    • 제38권7호
    • /
    • pp.624-629
    • /
    • 2015
  • Since the emergence of proteomics methods, many proteins specific for renal cell carcinoma (RCC) have been identified. Despite their usefulness for the specific diagnosis of RCC, such proteins do not provide spatial information on the diseased tissue. Therefore, the identification of cancer-specific proteins that include information on their specific location is needed. Recently, matrix-assisted laser desorption ionization (MALDI) mass spectrometry (MS) based imaging mass spectrometry (IMS) has emerged as a new tool for the analysis of spatial distribution as well as identification of either proteins or small molecules in tissues. In this report, surgical tissue sections of papillary RCC were analyzed using MALDI-IMS. Statistical analysis revealed several discriminative cancer-specific m/z-species between normal and diseased tissues. Among these m/z-species, two particular proteins, S100A11 and ferritin light chain, which are specific for papillary RCC cancer regions, were successfully identified using LC-MS/MS following protein extraction from independent RCC samples. The expressions of S100A11 and ferritin light chain were further validated by immunohistochemistry of human tissues and tissue microarrays (TMAs) of RCC. In conclusion, MALDI-IMS followed by LC-MS/MS analysis in human tissue identified that S100A11 and ferritin light chain are differentially expressed proteins in papillary RCC cancer regions.

Iron Starvation-Induced Proteomic Changes in Anabaena (Nostoc) sp. PCC 7120: Exploring Survival Strategy

  • Narayan, Om Prakash;Kumari, Nidhi;Rai, Lal Chand
    • Journal of Microbiology and Biotechnology
    • /
    • 제21권2호
    • /
    • pp.136-146
    • /
    • 2011
  • This study provides first-hand proteomic data on the survival strategy of Anabaena sp. PCC 7120 when subjected to long-term iron-starvation conditions. 2D-gel electrophoresis followed by MALDI-TOF/MS analysis of iron-deficient Anabaena revealed significant and reproducible alterations in ten proteins, of which six are associated with photosynthesis and respiration, three with the antioxidative defense system, and the last, hypothetical protein all1861, conceivably connected with iron homeostasis. Iron-starved Anabaena registered a reduction in growth, photosynthetic pigments, PSI, PSII, whole-chain electron transport, carbon and nitrogen fixation, and ATP and NADPH content. The kinetics of hypothetical protein all1861 expression, with no change in expression until day 3, maximum expression on the $7^{th}$ day, and a decline in expression from the $15^{th}$ day onward, coupled with in silico analysis, suggested its role in iron sequestration and homeostasis. Interestingly, the up-regulated FBP-aldolase, Mn/Fe-SOD, and all1861 all appear to assist the survival of Anabeana subjected to iron-starvation conditions. Furthermore, the $N_2$-fixation capabilities of the iron-starved Anabaena encourage us to recommend its application as a biofertilizer, particularly in iron-limited paddy soils.

Comparison of Plasma Proteome Expression between the Young and Mature Adult Pigs

  • Jeong, Jin Young;Nam, Jin Sun;Kim, Jang Mi;Jeong, Hak Jae;Kim, Kyung Woon;Lee, Hyun-Jeong
    • Reproductive and Developmental Biology
    • /
    • 제37권4호
    • /
    • pp.247-253
    • /
    • 2013
  • Here, we present an approach of blood plasma proteome profiling and their comparisons between the young and the adult pigs as prerequisite for the identification of bio-markers related to the health conditions, growth performance and meat quality. To profile the proteome in porcine plasma, blood samples were collected from 19 young piglets and 20 adult male barrows and the plasma was retrieved. Then, protein profiling was initiated using one and two-dimensional electrophoresis. Proteins were spotted and then identified by MALDI-TOF-TOF and LC-MS-MS. In the results, more than thirty-six and twenty eight protein spots were selected in young piglets and adult pigs, respectively and twenty three proteins were identified. The proteome profile images were compared between those ones using Image Master Version 7.0. The image of expressed proteome showed that most of proteins from plasma of young piglet separated clearly and concentrated in 2DE display compared to ones from adult. Image analysis in detail was carried out to look for the specific proteins related to age progression. It demonstrated that the characteristics of proteome expression could be distinct to their age stages. Further investigations needed to proceed to understand the age dependent change of protein conformation and biological meaning of those differences in proteome expression between young and mature adult pigs.

공기 중에서 금속 헥사카르보닐 착물 $M(CO)_6$ (M=Cr, Mo, W)를 이용한 폴러렌 산화물 $[C_{70}O_n](n=1{\sim}2)$의 초음파화학 합성 (Sonochemical Synthesis of Fullerene Oxides $[C_{70}O_n](n=1{\sim}2)$ Using Metal Hexacarbonyl Complexes $M(CO)_6$ (M=Cr, Mo, W) Under Air Atmosphere)

  • 고원배;박영환
    • Elastomers and Composites
    • /
    • 제40권3호
    • /
    • pp.174-180
    • /
    • 2005
  • 공기 중 초음파 조건에서 풀러렌$[C_{70}]$과 금속 헥사카르보닐 착물 $M(CO)_6$ (M=Cr, Mo, W)을 반응시켜 풀러렌 산화물 $[C_{70}O_n](n=1{\sim}2)$을 합성하였다. 동일한 초음파 조건에서 풀러렌$[C_{70}]$과 여러 가지 금속 헥사카르보닐 착물 $M(CO)_6$ (M=Cr, Mo, W)의 반응성은 $Mo(CO)_6$ > $W(CO)_6$ > $Cr(CO)_6$ 순으로 증가함을 나타냈다. MALDI-TOF-MS, UV-visible 스펙트럼과 HPLC를 사용하여 분석한 결과 초음파화학 반응의 생성물은 $[C_{70}O_n](n=1{\sim}2)$ 임을 알 수 있었다.

초음파 조건에서 여러가지 아민 N-옥시드와 풀러렌($C_{70}$)를 이용한 풀러렌 산화물의 합성 (Synthesis of Fullerene Oxide by Fullerene ($C_{70}$) and Various Amino N-Oxides under Ultrasonic Irradiation)

  • 고원배;한지연;임영아;신원직;유구용
    • Elastomers and Composites
    • /
    • 제39권2호
    • /
    • pp.153-160
    • /
    • 2004
  • 풀러렌($C_{70}$)을 3-picoline N-oxide, pyridine N-oxide hydrate, quinoline N-oxide, isoquinolne N-oxide의 여러가지 아민 N-옥시드를 사용하여 $25{\sim}43^{\circ}C$, 초음파 조건에서 반응시켜 풀러렌 산화물을 합성 하였다. 동일한 초음파 조건에서 여러가지 아민 N-옥시드에 대한 풀러렌 ($C_{70}$)의 반응성은 3-picoline N-oxide : pyridine N-oxide hydrate : quinoline N-oxide : isoquinoline N-oxide 처럼 동일하게 나타났다. MALDI -TOF MS, UV-vis 그리고 HPLC를 사용하여 생성된 풀러렌 산화물을 분석한 결과 [$C_{70}(O)n$] (n=1) 임을 알 수 있었다.

마이크로파 조건에서 여러가지 산화제를 이용한 풀러렌[$C_{60}$의 산화반응 (The Oxidation of Fullerene[$C_{60}$] using Several Oxidants under Microwave Irradiation)

  • 고원배;황성호;안주현
    • Elastomers and Composites
    • /
    • 제40권1호
    • /
    • pp.45-52
    • /
    • 2005
  • 풀러렌[$C_{60}$]은 3-chloroperoxy benzoic acid, benzoyl peroxide, trichloroisocyanuric acid, chromium(VI) oxide 등의 산화제를 사용하여 마이크로파 조건에서 반응시켜 풀러렌 산화물[$C_{60}(O)_n$] ($n=1{\sim}4$ or n=1)을 합성하였다. 동일한 마이크로파 조건에서 여러 가지 산화제와 풀러렌[$C_{60}$]의 고체상태 반응성은 3-chloroperoxy benzoic acid>benzoyl peroxide>trichloroisocyanuric acid$\simeq$chromium(VI) oxide 순으로 증가함을 나타냈다. MALDI-TOF-MS, UV-visible, 그리고 HPLC를 사용하여 분석한 결과 생성된 풀러렌 산화물은[$C_{60}(O)_n$] ($n=1{\sim}4$ or n=1)임을 알 수 있었다.

마이크로파 조건에서 풀러렌 산화물 [$C_{70}O_n$ ($n=1{\sim}3$ or n=1)의 합성 (Synthesis of Fullerene Oxides [$C_{70}O_n$] ($n=1{\sim}3$ or n=1) under Microwave Irradiation)

  • 고원배;안주현;임영아;한지연;한동설
    • Elastomers and Composites
    • /
    • 제39권4호
    • /
    • pp.309-317
    • /
    • 2004
  • 풀러렌[$C_{70}$]을 3-chloroperoxy benzoic acid, chromium (VI) oxide, benzoyl peroxide, trichloroisocyanuric acid 등의 산화제를 사용하여 마이크로파 조건에서 반응시켜 풀러렌 난화물[$C_{70}O_n$] ($n=1{\sim}3$ or n=1)을 합성하였다. 동일한 마이크로파 조건에서 여러가지 산화제와 풀러렌[$C_{70}$]의 고체상태 반응성은 3-chloroperoxy benzoic acid > chromium(VI) oxide > trichloroisocyanuric acid $\simeq$ benzoyl peroxide 순으로 증가함을 나타냈다. MALDI-TOF-MS, UV-visible, 그리고 HPLC를 사용하여 분석한 결과 생성된 풀러렌 산화물은 [$C_{70}O_n$] ($n=1{\sim}3$ or n=1)임을 알 수 있었다.

Mass-Spectral Identification of an Extracellular Protease from Bacillus subtilis KCCM 10257, a Producer of Antibacterial Peptide Subtilein

  • SONG HYUK-HWAN;GIL MI-JUNG;LEE CHAN
    • Journal of Microbiology and Biotechnology
    • /
    • 제15권5호
    • /
    • pp.1054-1059
    • /
    • 2005
  • An extracellular protease was identified from Bacillus subtilis KCCM 10257 by N-terminal sequencing and mass spectral analysis. The molecular mass of the extracellular protease was estimated to be 28 kDa by SDS-PAGE. Sequencing of the N-terminal of the protease revealed the sequence of A(G,S,R)QXVPYG(A)V(P,L)SQ. The N-terminal sequence exhibited close similarity to the sequence of other proteases from Bacillus sp. A mass list of the monoisotopic peaks in the MALDI-TOF spectrum was searched after peptide fragmentation of the protease. Six peptide sequences exhibiting monoisotopic masses of 1,276.61, 1,513.67, 1,652.81, 1,661.83, 1,252.61, and 1,033.46 were observed from the fragmented protease. These monisotopic masses corresponded to the lytic enzyme L27 from Bacillus subtilis 168, and the Mowse score was found to be 75. A doubly charged Top product (MS) at a m/z of 517.3 exhibiting a molecular mass of 1034.6 was further analyzed by de novo sequencing using a PE Sciex QSTAR Hybrid Quadropole-TOF (MS/MS) mass spectrometer. MS/MS spectra of the Top product (MS) at a m/z of 517.3 obtained from the fragmented peptide mixture of protease with Q-star contained the b-ion series of 114.2, 171.2, 286.2, 357.2, 504.2, 667.4, 830.1, and 887.1 and y-ion series of 147.5, 204.2, 367.2, 530.3, 677.4, 748.4, 863.4, and 920.5. The sequence of analyzed peptide ion was identified as LGDAFYYG from the b- and y-ion series by de novo sequencing and corresponded to the results from the MALDI-TOF spectrum. From these results the extracellular protease from Bacillus subtilis KCCM 10257 was successfully identified with the lytic enzyme L27 from Bacillus subtilis 168.

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
    • /
    • 제13권6호
    • /
    • pp.859-865
    • /
    • 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.

Effect of Dihydroxybenzoic Acid Isomers on the Analysis of Polyethylene Glycols in MALDI-MS

  • Lee, Ae-Ra;Yang, Hyo-Jik;Kim, Yang-Sun;Kim, Jeong-Kwon
    • Bulletin of the Korean Chemical Society
    • /
    • 제30권5호
    • /
    • pp.1127-1130
    • /
    • 2009
  • The effects of different dihydroxybenzoic acid (DHB) isomers, when used as matrix materials in matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS), were investigated in analyses of polyethylene glycol (PEG) polymers. PEG polymers ranging from 400 to 8,000 Da were prepared in different DHB isomer matrices using solvent-based and solvent-free methods. PEG samples were detected only in matrices of 2,3-DHB, 2,5-DHB, and 2,6-DHB while the most intense peaks were observed using 2,6-DHB in both solvent-free and solvent-based preparations.