• Title/Summary/Keyword: p47

Search Result 7,087, Processing Time 0.03 seconds

Change in the Conformation of $p47^{phox}$ by Sodium Dodecyl Sulfate, an Activator of the Leukocyte NADPH Oxidase

  • Park, Jeen-Woo;Park, Hee-Sae
    • BMB Reports
    • /
    • v.31 no.3
    • /
    • pp.227-232
    • /
    • 1998
  • The leukocyte NADPH oxidase of neutrophils is a membrane-bound enzyme that catalyzes the production of $O_2^-$ from oxygen using NADPH as an electron donor. Dormant in resting neutrophils, the enzyme acquires catalytic activity when the cells are exposed to appropriate stimuli. During activation, the cytosolic oxidase components $p47^{phox}$ and $p67^{phox}$ migrate to the plasma membrane, where they associate with cytochrome $b_{558}$, a membrane-bound flavohemoprotein, to assemble the active oxidase. The oxidase can be activated in a cell-free system; the activating agent usually employed is an anionic amphiphile such as sodium dodecyl sulfate (SDS). Because $p47^{phox}$ can translocate by itself during activation, the conformational change in $p47^{phox}$ may be responsible for the activation of NADPH oxidase. We show here that the treatment of $p47^{phox}$ with SDS leads to an increase in the reactivity of the sutbydryl group of cysteines toward N-ethylmaleimide, indicating that the conformational change occurs when $p47^{phox}$ is exposed to SDS. We propose that this change in conformation results in the appearance of a binding site through which $p47^{phox}$ interacts with cytochrome $b_{558}$during the activation process.

  • PDF

Crystal Structure of p97 N-D1 Hexamer in Complex with p47 UBX Domain

  • Thang Quyet Nguyen;Wonchull Kang
    • Journal of the Korean Chemical Society
    • /
    • v.68 no.1
    • /
    • pp.25-31
    • /
    • 2024
  • The p97 adenosine triphosphatase is a key player in protein homeostasis, responsible for unfolding ubiquitylated substrates. It engages with various adaptor proteins through its N-terminal domain, with the p97-p47 complex attracting particular attention for its involvement in membrane remodeling. Although the structures of p97 in complex with the Ubiquitin regulatory X (UBX) domain from various adaptors have been reported, the stoichiometry is conflicting. Here, we report the crystal structure of the p97 N-D1 hexamer in complex with the p47 UBX domain at a resolution of 2.7 Å. The structure reveals a stoichiometry of 6:6 between the p97 N-D1 and the p47 UBX domain. These findings provide valuable insights into the binding stoichiometry of p97 N-D1 and p47 UBX domain, which are crucial for understanding the role of p97 and adaptor proteins in cellular processes such as the ubiquitin-proteasome pathway, membrane fusion, and cell cycle regulation.

Molecular cloning and expression of black rockfish Sebastes schlegelii p47-phox (neutrophil cytosolic factor 1)

  • Kim, Ki-Hyuk;Baeck, Gun-Wook;Kim, Mu-Chan;Park, Chan-Il
    • Journal of fish pathology
    • /
    • v.22 no.2
    • /
    • pp.137-146
    • /
    • 2009
  • The black rockfish Sebastes schlegelii neutrophil cytosolic factor components p47 phox (phagocyte oxidase) cDNA was cloned. The sequence of the cDNA showed that rockfish p47 phox cDNA consisted of 1,952 bp contained open reading frame encoding predicted polypeptide of 420 amino acids. Additionally analysis of the p47 phox amino acid sequence showed two potential SH3 domains. The functional domains are highly conserved in many animals, though the sequence of the components of the black rockfish showed low homology with that of mammals. The deduced amino acid sequence of the black rockfish p47 phox was similar to those of the carp (60.4%), zebrafish (59,2%), rainbow trout (68.5%), xenopus (55.2%), mouse (54.2%), rabbit (54.5%), rat (53.7%), and chicken (50.9%). The expression of the rockfish p47 phox molecule was induced in peripheral blood leukocytes (PBLs) from 1 to 12 h following LPS stimulation, with a peak at 6 h after the stimulation, and which increased at 1, 3, and 12 h after treated with Poly I:C compared with the control. The rockfish p47 phox gene was expressed in various tissues of healthy fish. The level of p47 phox expression was high in the PBLs, kidney and spleen.

Effect of Arginine Modification of Cytosolic Component $p47^{phox}$ by Phenylglyoxal on the Activation of Respiratory Burst Oxidase in Human Neutrophils

  • Park, Jeen-Woo
    • BMB Reports
    • /
    • v.29 no.6
    • /
    • pp.507-512
    • /
    • 1996
  • The NADPH oxidase of phagocytes catalyzes the reduction of oxygen to $O_{2}^{-}$ at the expense of NADPH The enzyme is dormant in resting neutrophils and hecomes activated on stimulation. During activation. $p47^{phox}$ (phagocyte oxidase factor), a cytosolic oxidase subunit, becomes extensively phosphorylated on a number of serines located between S303-S379. Although the biochemical role of phosphorylation is speculative, it has been suggested that phosphorylation could neutralize the strongly cationic C-terminal which may result in the change of conformation of $p47^{phox}$ and subsequent translocation of this protein and other cytosolic components to the membrane. In order to mimic the effect of phosphorylation in terms of neutralizing the positive charges, recombinant $p47^{phox}$ was treated with phenylglyoxal, which removes positive charges of arginine residues. Modification of recombinant $p47^{phox}$ resulted in the activation of oxidase in a cell-free translocation system as well as a conformational change in recombinant $p47^{phox}$ which may be responsible for the activation of the enzyme.

  • PDF

Nucleotide Binding Component of the Respiratory Burst Oxidase of Human Neutrophils

  • Park, Jeen-Woo;Ahn, Soo-Mi
    • BMB Reports
    • /
    • v.28 no.3
    • /
    • pp.275-280
    • /
    • 1995
  • The respiratory burst oxidase of neutrophils is a multicomponent enzyme, domant in resting cells, that catalyzes the reduction of oxygen to $O_{2}^{-}$ at the expense of NADPH. In the resting neutrophil, some of the components of the oxidase, including proteins p47 and p67, are in the cytosol, while the rest are in the plasma membrane. Recent evidence has suggested that at least some of the cytosolic oxidase components exist as a complex. The cytosolic complex with a molecular weight of ~240 kDa was found to bind to blue-agarose and 2',5'-ADP-agarose, which recognize nucleotide requiring enzymes. In order to identify the nucleotide binding component of the cytosolic complex we purified recombinant p47 and p67 fusion proteins using the pGEX system. Pure recombinant p47 was retained completely on 2',5'-ADP-agarose, whereas pure recombinant p67 did not bind to these affinity beads. On the basis of these results, we infer that p47 may contain the nucleotide binding site.

  • PDF

Comparative Evaluation of Silibinin Effects on Cell Cycling and Apoptosis in Human Breast Cancer MCF-7 and T47D Cell Lines

  • Jahanafrooz, Zohreh;Motameh, Nasrin;Bakhshandeh, Behnaz
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.17 no.5
    • /
    • pp.2661-2665
    • /
    • 2016
  • Silibinin is a natural polyphenol with high antioxidant and anticancer properties. In this study, its influence on two of the most commonly employed human breast cancer cell lines, MCF-7 and T47D, and one non-malignant MCF-10A cell line, were investigated and compared. Cell viability, the cell cycle distribution and apoptosis induction were analyzed by MTT and flow cytometry, respectively. The effect of silibinin on PTEN, Bcl-2, P21, and P27 mRNAs expression was also investigated by real-time RT-PCR. It was found that silibinin caused G1 cell cycle arrest in MCF-7 and MCF-10A cells but had no effect on the T47D cell cycle. Silibinin induced cytotoxic and apoptotic effects in T47D cells more than the MCF-7 cells and had no cytotoxic effect in MCF-10A cells under the same conditions. Silibinin upregulated PTEN in MCF-7 and caused slightly increased P21 mRNA expression in T47D cells and slightly increased PTEN and P21 expression in MCF-10A cells. Bcl-2 expression decreased in all of the examined cells under silibinin treatment. P27 mRNA expression upregulated in T47D and MCF-10A cells under silibinin treatment. PTEN mRNA in T47D and P21 and P27 mRNAsin MCF-7 were not affected by silibinin. These results suggest that silibinin has mostly different inhibitory effects in breast cancer cells and might be an effective anticancer agent for some cells linked to influence on cell cycle progression.

Physicochemical Properties of a Giant Embryo Mutant Induced by T-DNA Insertion in Rice

  • Park, Hee-Yeon;Qin, Yang;Sohn, Jae-Keun
    • KOREAN JOURNAL OF CROP SCIENCE
    • /
    • v.56 no.4
    • /
    • pp.413-419
    • /
    • 2011
  • This study was conducted to determine the physicochemical properties of a giant embryo rice 'P47JB-4-B-5-B' derived from the cross between 'P47', a mutant of 'Hwayoung' induced by T-DNA insertion, and 'Junam'. The grain appearance and chemical components of the embryo were analyzed and compared with a donor cultivar, 'Hwayoung'. The proportion of embryo weight to grain weight of 'P47 JB-4-B-5-B' was 2.2 times heavier (6.7%) than that (3.1%) of 'Hwayoung'. Total free amino acid content (75.81 mg/100 g) of 'P47JB-4-B-5-B' was 2.1 times higher than that of 'Hwayoung'. The GABA content in brown rice was 14.06 mg/100 g in 'P47JB-4-B-5-B' and 6.8 mg/100 g in 'Hwayoung'. Especially, the GABA content in brown rice of 'P47JB-4-B-5-B' remarkably increased (about 33 times from 1.48 mg to 44.81 mg/100 g) 2 days after germination. Continuous frequency distributions and transgressive segregation in embryo length and width were observed in the $F_2$ population of the cross between 'P47' and 'Cheongcheong', indicating that the giant embryo was controlled by quantitative trait loci. However, embryo length and width demonstrated high broad sense heritability, implying that giant embryonic traits could be selected in earlier generations in comparison with other quantitative traits.

Effect of Mindful Self-Compassion Training on Anxiety, Depression and Emotion Regulation

  • Kim, Sunghwan;Song, Yungsook;Lee, Kyoung-Uk
    • Anxiety and mood
    • /
    • v.18 no.1
    • /
    • pp.10-16
    • /
    • 2022
  • Objective : The present study examined the effectiveness of the mindful self-compassion (MSC) program on anxiety, depression, self-compassion and emotion regulation. Methods : A total of 29 subjects (mean age 27.5±6.5 years, 15 males and 14 females) participated in a standardized 8-week MSC program. The control group consisted of age- and sex-matched twenty participants (mean age 26.0±2.9 years, 11 males and 9 females). All subjects completed self-report measurements at two weeks before and after the MSC program. Results : MSC training improved self-compassion as demonstrated by the significant group x time interaction effects on the total Self-Compassion Scale scores (F[1, 47]=8.324, p<0.01). Regarding the subscale scores, a significant improvement in self-kindness, isolation and mindfulness components of self-compassion was observed after MSC training. A significant group x time interaction was observed on the self-kindness subscale (F[1, 47]=4.664, p<0.05), with a significant main effect of time (F[1, 47]=23.723, p<0.001). The isolation subscale showed a significant group x time interaction (F[1, 47]=8.698, p<0.001). For the mindfulness subscale, there was a significant group x time interaction (F[1, 47]=6.611, p<0.05) and main effect of time (F[1, 47]=6.611, p<0.05). MSC training also improved the acceptance emotion regulation strategy, as demonstrated by the significant group x time interaction in the acceptance subscale scores of the Cognitive Emotion Regulation Questionnaire (F[1, 47]=6.845, p<0.05). Conclusion : MSC training showed efficacy in fostering self-compassion and improving emotion regulation. Thus, this program might be applicable to improve mental health.

Biodegradation of Cutting Oil by Pseudomonas aeruginosa KS47 (Pseudomonas aeruginosa KS47에 의한 절삭유의 생물학적 분해)

  • Kim, Lan-Hee;Lee, Sang-Seob
    • Korean Journal of Microbiology
    • /
    • v.44 no.1
    • /
    • pp.22-28
    • /
    • 2008
  • Cutting oils are emulsionable fluids widely used in metal working processes. Their composition is mineral oil, water, and additives (fatty acids, surfactants, biocides, etc.) generating a toxic waste after a long use. Cutting oils also affect colour, taste and odour of water, making it undesirable for domestic and industrial uses. In these days, conventional treatment methods as evaporation, membrane separation or chemical separation have major disadvantages since they generate a concentrated stream that is more harmful than the original waste. In this study, our purpose is to reduce cutting oils by using biological treatment. Eighty one strains were isolated from cutting waste oil of industrial waste water sludge under aerobic conditions. Among these strains, KS47, which removed 90.4% cutting oil in 48 hr, was obtained by screening test under aerobic conditions(pH 7, $28^{\circ}C$). KS47 was identified as Pseudomonas aeruginosa according to morphological, physiological and biochemical properties, 16S rDNA sequence, and fatty acid analysis. P. aeruginosa KS47 could utilize cutting oil as carbon source. In batch test, we obtained optimal degradation conditions(1.5 g/L cell concentration, pH 7, and temperature $30^{\circ}C$). Under the optimal conditions, 1,060 mg/L cutting oil was removed 83.7% (74.1 mg/L/hr).

Genetic Structure of xyl Gene Cluster Responsible for Complete Degradation of (4-Chloro )Benzoate from Pseudomonas sp. S-47

  • Park, Dong-Woo;Lee, Kyoung;Chae, Jong-Chan;Kudo, Toshiaki;Kim, Chi-Kyung
    • Journal of Microbiology and Biotechnology
    • /
    • v.14 no.3
    • /
    • pp.483-489
    • /
    • 2004
  • Pseudomonas sp. S-47 is a bacterium capable of degrading benzoate as well as 4-chlorobenzoate (4CBA). Benzoate and 4CBA are known to be degraded via a meta-cleavage pathway characterized by a series of enzymes encoded by xyl genes. The meta-cleavage pathway operon in Pseudomonas sp. S-47 encodes a set of enzymes which transform benzoate and 4CBA into TCA cycle intermediates via the meta-cleavage of (4-chloro )catechol to produce pyruvate and acetyl-CoA. In the current study, the meta-pathway gene cluster was cloned from the chromosomal DNA of S-47 strain to obtain pCS1, which included the degradation activities for 4CBA and catechol. The genetic organization of the operon was then examined by cloning the meta-pathway genes into a pBluescript SKII(+) vector. As such, the meta-pathway operon from Pseudomonas sp. S-47 was found to contain 13 genes in the order of xylXYZLTEGFlQKIH. The two regulatory genes, xylS and xylR, that control the expression of the meta-pathway operon, were located adjacently downstream of the meta-pathway operon. The xyl genes from strain S-47 exhibited a high nucleoside sequence homology to those from Pseudomonas putida mt-2, except for the xylJQK genes, which were more homologous to the corresponding three genes from P. stutzeri AN10. One open reading frame was found between the xylH and xylS genes, which may playa role of a transposase. Accordingly, the current results suggest that the xyl gene cluster in Pseudomonas sp. S-47 responsible for the complete degradation of benzoate was recombined with the corresponding genes from P. putida mt-2 and P. stutzeri AN10.