• Title/Summary/Keyword: molecular modification

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Changes in Protein Phosphorylation during Salivary Gland Degeneration in Haemaphysalis longicornis

  • Xiao, Qi;Hu, Yuhong;Yang, Xiaohong;Tang, Jianna;Wang, Xiaoshuang;Xue, Xiaomin;Li, Mengxue;Wang, Minjing;Zhao, Yinan;Liu, Jingze;Wang, Hui
    • Parasites, Hosts and Diseases
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    • v.58 no.2
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    • pp.161-171
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    • 2020
  • The ticks feed large amount of blood from their hosts and transmit pathogens to the victims. The salivary gland plays an important role in the blood feeding. When the female ticks are near engorgement, the salivary gland gradually loses its functions and begins to rapidly degenerate. In this study, data-independent acquisition quantitative proteomics was used to study changes in the phosphorylation modification of proteins during salivary gland degeneration in Haemaphysalis longicornis. In this quantitative study, 400 phosphorylated proteins and 850 phosphorylation modification sites were identified. Trough RNA interference experiments, we found that among the proteins with changes in phosphorylation, apoptosis-promoting Hippo protein played a role in salivary gland degeneration.

Molecular Cloning and Characterization of Bacillus cereus O-Methyltransferase

  • Lee Hyo-Jung;Kim Bong-Gyu;Ahn Joong-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.16 no.4
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    • pp.619-622
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    • 2006
  • Biotransformation is a good tool to synthesize regioselective compounds. It could be performed with diverse sources of genes, and microorganisms provide a myriad of gene sources for biotransformation. We were interested in modification of flavonoids, and therefore, we cloned a putative O-methyltransferase from Bacillus cereus, BcOMT-2. It has a 668-bp open reading frame that encodes a 24.6-kDa protein. In order to investigate the modification reaction mediated by BcOMT-2, it was expressed in E. coli as a His-tag fusion protein and purified to homogeneity. Several substrates such as naringenin, luteolin, kaempferol, and quercetin were tested and reaction products were analyzed by thin layer chromatography (TLC) and high performance liquid chromatography (HPLC). BcOMT-2 could transfer a methyl group to substrates that have a 3' functional hydroxyl group, such as luteolin and quercetin. Comparison of the HPLC retention time and UV spectrum of the quercetin reaction product with corresponding authentic 3'-methylated and 4'-methylated compounds showed that the methylation position was at either the 3'-hydroxyl or 4'-hydroxyl group. Thus, BcOMT-2 transfers a methyl group either to the 3'-hydroxyl or 4'-hydroxyl group of flavonoids when both hydroxyl groups are available. Among several flavonoids that contain a 3'- and 4'-hydroxyl group, fisetin was the best substrate for the BcOMT-2.

The role of protein arginine-methyltransferase 1 in gliomagenesis

  • Wang, Shan;Tan, Xiaochao;Yang, Bin;Yin, Bin;Yuan, Jiangang;Qiang, Boqin;Peng, Xiaozhong
    • BMB Reports
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    • v.45 no.8
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    • pp.470-475
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    • 2012
  • Protein arginine methyltransferase 1 (PRMT1), a type-I arginine methyltransferase, has been implicated in diverse cellular events. We have focused on the role of PRMT1 in gliomagenesis. In this study, we showed that PRMT1 expression was up-regulated in glioma tissues and cell lines compared with normal brain tissues. The knock-down of PRMT1 resulted in an arrest in the G1-S phase of the cell cycle, proliferation inhibition and apoptosis induction in four glioma cell lines (T98G, U87MG, U251, and A172). Moreover, an in vivo study confirmed that the tumor growth in nude mouse xenografts was significantly decreased in the RNAi-PRMT1 group. Additionally, we found that the level of the asymmetric dimethylated modification of H4R3, a substrate of PRMT1, was higher in glioma cells than in normal brain tissues and decreased after PRMT1 knock-down. Our data suggest a potential role for PRMT1 as a novel biomarker of and therapeutic target in gliomas.

Modification and inactivation of Cu,Zn-superoxide dismutase by the lipid peroxidation product, acrolein

  • Kang, Jung Hoon
    • BMB Reports
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    • v.46 no.11
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    • pp.555-560
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    • 2013
  • Acrolein is the most reactive aldehydic product of lipid peroxidation and is found to be elevated in the brain when oxidative stress is high. The effects of acrolein on the structure and function of human Cu,Zn-superoxide dismutase (SOD) were examined. When Cu,Zn-SOD was incubated with acrolein, the covalent crosslinking of the protein was increased, and the loss of enzymatic activity was increased in a dose-dependent manner. Reactive oxygen species (ROS) scavengers and copper chelators inhibited the acrolein-mediated Cu,Zn-SOD modification and the formation of carbonyl compound. The present study shows that ROS may play a critical role in acrolein-induced Cu,Zn-SOD modification and inactivation. When Cu,Zn-SOD that has been exposed to acrolein was subsequently analyzed by amino acid analysis, serine, histidine, arginine, threonine and lysine residues were particularly sensitive. It is suggested that the modification and inactivation of Cu,Zn-SOD by acrolein could be produced by more oxidative cell environments.

Chemical Modification of Lysine Residues in Bacillus licheniformis α-Amylase: Conversion of an Endo- to an Exo-type Enzyme

  • Habibi, Azadeh Ebrahim;Khajeh, Khosro;Nemat-Gorgani, Mohsen
    • BMB Reports
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    • v.37 no.6
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    • pp.642-647
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    • 2004
  • The lysine residues of Bacillus licheniformis $\alpha$-amylase (BLA) were chemically modified using citraconic anhydride or succinic anhydride. Modification caused fundamental changes in the enzymes specificity, as indicated by a dramatic increase in maltosidase and a reduction in amylase activity. These changes in substrate specificity were found to coincide with a change in the cleavage pattern of the substrates and with a conversion of the native endo- form of the enzyme to a modified exo- form. Progressive increases in the productions of $\rho$-nitrophenol or glucose, when para nitrophenyl-maltoheptaoside or soluble starch, respectively, was used as substrate, were observed upon modification. The described changes were affected by the size of incorporated modified reagent: citraconic anhydride was more effective than succinic anhydride. Reasons for the observed changes are discussed and reasons for the effectivenesses of chemical modifications for tailoring enzyme specificities are suggested.

Micro/Nano Adhesion and Friction Characteristics of PTFE Coating Film Deposited by IBAD Method (IBAD 방법으로 코팅된 PTFE 박막의 마이크로/나노 응착 및 마찰 특성)

  • 윤의성;오현진;한흥구;공호성;장경영
    • Tribology and Lubricants
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    • v.20 no.5
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    • pp.237-244
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    • 2004
  • Micro/nano tribological characteristics of PTFE coating films were experimentally studied. PTFE (polytetrafluoroethylene) modified polyethylene and low molecular weight PTFE were used as a coating materials. These films were deposited on Si-wafer (100) by IBAD (ion beam assisted deposition) method. The Ar ion beam sputtering was performed to change the surface topography of films using a hollow cathode ion gun under different Ar ion dose conditions in a vacuum chamber. Micro/nano tribological characteristics, water wetting angles and roughness were measured with a micro tribotester, SPM (scanning probe microscope), contact anglemeter and profilometer, respectively. The durability of the films were measured with macro tribotester. Results showed that the PTFE coating surfaces were converted to hydrophobic. The water contact angle of coated surfaces and surface roughness increased with the coating thickness. Adhesion and friction in micro and nano scale were governed by magnitude of normal load in soft material such as PTFE films. As the increase of sputtering time on low molecular weight PTFE films, the surface roughness was increased and nano adhesion and friction were decreased. The nano tribological characteristics of surfaces are mainly improved by chemical modification such as PTFE coating and given a synergy effect by the physical modification such as topographic modification.