Hypoxia‐induced expression of phosphoglycerate mutase B in fibroblasts

Takahashi, Yûji; Takahashi, Shigeru; Yoshimi, Tatsuya; Miura, Takashi

doi:10.1046/j.1432-1327.1998.2540497.x

Cited by 34 publications

(28 citation statements)

References 9 publications

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“…Although PGAM1 may contribute to an increase in glycolytic flux in response to hypoxia, PGAM1 levels increasing in fibroblasts cultured in hypoxic conditions (31), increased glycolysis has also been associated with protection against oxidative stress (15). PGAM1 has several phosphorylation sites.…”

Section: Discussionmentioning

confidence: 99%

Proteomic analysis of epicardial and subcutaneous adipose tissue reveals differences in proteins involved in oxidative stress

Salgado-Somoza¹,

Teijeira-Fernández

Fernández

et al. 2010

American Journal of Physiology-Heart and Circulatory Physiology

144

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Salgado-Somoza A, Teijeira-Fernández E, Fernández AL, González-Juanatey JR, Eiras S. Proteomic analysis of epicardial and subcutaneous adipose tissue reveals differences in proteins involved in oxidative stress. Am J Physiol Heart Circ Physiol 299: H202-H209, 2010. First published April 30, 2010; doi:10.1152/ajpheart.00120.2010.-Epicardial adipose tissue (EAT) is an endocrine organ adjacent to coronary arteries and myocardium without anatomy barriers. Locally produced adipokines may reflect or affect to cardiovascular physiology and pathology. Our aim was to study the protein expression profiles of EAT and subcutaneous adipose tissue (SAT) to identify local candidate molecules characterizing EAT in patients with cardiovascular disease. EAT and SAT samples were collected from 55 patients undergoing heart surgery. Proteins from these tissues were separated by two-dimensional (2D) gel electrophoresis, and differences between them were identified by MALDI-TOF/TOF spectra. Differences in protein levels were further investigated by real-time RT-PCR and Western blots, and production of reactive oxygen species (ROS) in EAT and SAT was evaluated by nitroblue tetrazolium chloride assays. ROS production was higher in EAT than SAT. We have found mRNA differences for catalase, glutathione S-transferase P, and protein disulfide isomerase, and 2D Western blots additionally showed post-translational differences for phosphoglycerate mutase 1; all four are related to oxidative stress pathways. EAT suffers greater oxidative stress than SAT in patients with cardiovascular diseases and exhibits associated proteomic differences that suggest the possibility of its association with myocardial stress in these patients. epicardial adipose tisssue OBESITY IS ONE OF THE MAIN causes of metabolic diseases. It is associated with a chronic inflammatory response and cardiovascular diseases (34). However, risk depends significantly on the distribution of adipose tissue in the body; for example, metabolic risk factors were more associated with omental adipose tissue (OAT) than with subcutaneous adipose tissue (SAT) among 3,001 participants in the Framingham Heart Study (6). These differences are plausibly due to different adipose tissues differing in adipokine expression and the production of inflammatory reactive oxygen species (ROS), which are known to be associated with cardiovascular disease (8); OAT is associated with more circulating substances related to inflammation and oxidative stress than SAT is (24).Epicardial adipose tissue (EAT) is a visceral fat pad adjacent to coronary arteries and the myocardium (11). The amount of EAT is related to left ventricle mass (13) and to the amount of intra-abdominal visceral fat, and visceral adiposity can in fact be evaluated by means of its echocardiographic measurement (10). Like other adipose tissues, EAT is now recognized as an endocrine organ, and its proximity to myocardium and coronary arteries suggests the possibility of paracrine action on these structures. In fact, in patients with coronary ...

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Section: Discussionmentioning

confidence: 99%

Proteomic analysis of epicardial and subcutaneous adipose tissue reveals differences in proteins involved in oxidative stress

Salgado-Somoza¹,

Teijeira-Fernández

Fernández

et al. 2010

American Journal of Physiology-Heart and Circulatory Physiology

144

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“…At subconfluence, cells were exposed to hypoxia or treated with chemicals. Exposures of cells to hypoxia were performed as described previously (9). In some experiments, 200 M ascorbic acid was added to the medium.…”

Section: Methodsmentioning

confidence: 99%

“…Differential Display and Transcriptional Run-on Assay-We followed the fluorescent differential display method developed by Ito et al (20) with several modifications, as described in a previous paper (9). Transcriptional run-on assay was performed as described previously (9).…”

Section: Methodsmentioning

confidence: 99%

Hypoxic Induction of Prolyl 4-Hydroxylase α(I) in Cultured Cells

Takahashi¹,

Takahashi²,

Shiga

et al. 2000

Journal of Biological Chemistry

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160

102

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Accumulated evidence indicates that hypoxia activates collagen synthesis in tissues. To explore the molecular mechanism of activation, we screened genes that are up-regulated or down-regulated by hypoxia. Fibroblasts isolated from fetal rat lung were cultured under hypoxia. Differential display technique showed that the mRNA level of prolyl 4-hydroxylase (PH) ␣(I), an active subunit that catalyzes the oxygen-dependent hydroxylation of proline residue in procollagen, increased 2-3-fold after an 8-h exposure to hypoxia. This elevated level was maintained over 40 h and returned to the basal level after reoxygenation. The transcription rate, protein level, and hydroxyproline content (an indicator of the prolyl hydroxylation) were all elevated by hypoxic culture. Analysis of the promotor region of PH␣(I) gene indicated that a motif similar to hypoxia-responsive element (HRE) of hypoxia-inducible genes such as erythropoietin, was identified within a 120-base pair sequence upstream of the transcription start site. Luciferase reporter assay and mutational analysis showed that a site similar to the HRE in this motif is functionally essential to hypoxic response. Electrophoretic mobility shift assay revealed that hypoxia-inducible factor-1 was stimulated and bound to the PH␣(I) HRE upon hypoxic challenge. Our results indicate that PH␣(I), an essential enzyme for collagen synthesis, is a target gene for hypoxia-inducible factor-1.Restricted oxygen availability is a feature of many physiologic and pathologic conditions, including high altitude residence, fetal development in the uterus, pulmonary fibrosis, wounded tissue, and neoplasm (1). Systemic and cellular responses to reduced oxygen tension (hypoxia) are initiated by activation and/or inactivation of gene expression. Hypoxia-inducible factor-1 (HIF-1), 1 which was originally found to be a critical mediator for the inducible expression of the erythropoietin (Epo) gene by hypoxia (2), is a heterodimer composed of HIF-1␣ and arylhydrocarbon receptor nuclear translocator (ARNT). HIF-1␣ and ARNT retain a basic helix-loop-helix domain and a Per-ARNT/aryl hydrocarbon receptor Sim domain in their N termini (2). Hypoxia induces stabilization of HIF-1␣ (3), heterodimerization of HIF-1␣ and ARNT (4), and the binding of the heterodimer to the hypoxia-responsive element (HRE) in the regulatory region of the target genes with the transcriptional coactivator p300/CREB-binding protein (5). Although posttranscriptional mechanisms may contribute to the induction of hypoxia-sensitive genes, activation of the HIF-1 complex is an important step leading to hypoxia-mediated induction of glycolytic enzymes (6 -9), Epo (2), vascular endothelial growth factor (10), and tyrosine hydroxylase (11).In the remodeling of the small muscular pulmonary artery observed in hypoxia-induced pulmonary hypertension, type I collagen is actively synthesized and accumulated in the media and the adventitia of the artery (12). Recent studies have revealed that in vivo exposure of rats to hypoxia increases prolyl ...

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“…3B). Edman degradation sequence analysis identified three N-terminal sequences: 30 FEIALP, 369 KFVPGC, and 370 FVPGC. The first of these has previously been reported by us as the amino-terminal of 135 kDa CDCP1 (2), generated through removal of the 29 residue signal peptide (Fig.…”

Section: Post-translational Processing Of 135-kda Cdcp1mentioning

confidence: 99%

Proteolysis-induced N-terminal Ectodomain Shedding of the Integral Membrane Glycoprotein CUB Domain-containing Protein 1 (CDCP1) Is Accompanied by Tyrosine Phosphorylation of Its C-terminal Domain and Recruitment of Src and PKCδ

Wortmann

Burke

et al. 2010

Journal of Biological Chemistry

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CUB-domain-containing protein 1 (CDCP1) is an integral membrane glycoprotein with potential as a marker and therapeutic target for a number of cancers. Here we examine mechanisms regulating cellular processing of CDCP1. By analyzing cell lines exclusively passaged non-enzymatically and through use of a panel of protease inhibitors, we demonstrate that full-length 135 kDa CDCP1 is post-translationally processed in a range of cell lines by a mechanism involving serine protease activity, generating a C-terminal 70-kDa fragment. Immunopurification and N-terminal sequencing of this cell-retained fragment and detailed mutagenesis, show that proteolytic processing of CDCP1 occurs at two sites, Arg-368 and Lys-369. We show that the serine protease matriptase is an efficient, but not essential, cellular processor of CDCP1 at Arg-368. Importantly, we also demonstrate that proteolysis induces tyrosine phosphorylation of 70-kDa CDCP1 and recruitment of Src and PKC␦ to this fragment. In addition, Western blot and mass spectroscopy analyses show that an N-terminal 65-kDa CDCP1 ectodomain is shed intact from the cell surface. These data provide new insights into mechanisms regulating CDCP1 and suggest that the biological role of this protein and, potentially, its function in cancer, may be mediated by both 70-kDa cell retained and 65-kDa shed fragments, as well as the full-length 135-kDa protein. CUB-domain-containing protein 1 (CDCP1)3 is an 836 amino acid integral membrane glycoprotein with a type I orientation at the cell surface (1-4), that is up-regulated in a number of malignancies including breast (1, 5, 6), colon (1, 2, 7), and lung (1) cancers. Of potential clinical significance, CDCP1 expression correlates with recurrence and patient survival rate in renal cell carcinomas (8) and lung adenocarcinomas (9), indicating that it may be suitable as a prognostic marker. Consistent with a role in cancer progression, silencing of CDCP1 reduced the metastatic ability of lung cancer A549 cells (10) and the peritoneal dissemination of gastric cancer 44As3 cells (11) in mice. Although its biological function is not known, the potential of CDCP1 as a therapeutic target for cancer treatment has been highlighted by studies showing that antibody-mediated inhibition of CDCP1 reduced metastasis of prostate cancer PC3 cells in mice (12, 13) and chicken embryos (13). Currently the mechanisms regulating CDCP1 in cancer and normal physiology are not well defined (14).During cellular processing, the 29-residue CDCP1 N-terminal signal peptide is removed generating a protein with molecular mass identified as either 135 kDa (2, 10, 15) or 140 kDa (3, 4) that contains 30 -40 kDa of N-linked glycans (2). In addition to this full-length form, there is evidence that a shorter CDCP1 species is expressed endogenously by a range of cell lines or is generated through the action of exogenous serine proteases. For example, lung cancer A549, PC14, H520, H322, and H157 cells (10) and gastric cancer 44As3 and 58As9 cells (11) resuspended non-enzymatically ...

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Hypoxia‐induced expression of phosphoglycerate mutase B in fibroblasts

Cited by 34 publications

References 9 publications

Proteomic analysis of epicardial and subcutaneous adipose tissue reveals differences in proteins involved in oxidative stress

Proteomic analysis of epicardial and subcutaneous adipose tissue reveals differences in proteins involved in oxidative stress

Hypoxic Induction of Prolyl 4-Hydroxylase α(I) in Cultured Cells

Proteolysis-induced N-terminal Ectodomain Shedding of the Integral Membrane Glycoprotein CUB Domain-containing Protein 1 (CDCP1) Is Accompanied by Tyrosine Phosphorylation of Its C-terminal Domain and Recruitment of Src and PKCδ

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