排序方式: 共有13条查询结果,搜索用时 31 毫秒
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Annelize Z. B. Arag?o Maria Luiza C. Nogueira Daniela C. Granato Fernando M. Simabuco Rodrigo V. Honorato Zaira Hoffman Sami Yokoo Francisco R. M. Laurindo Fabio M. Squina Ana Carolina M. Zeri Paulo S. L. Oliveira Nicholas E. Sherman Adriana F. Paes Leme 《The Journal of biological chemistry》2012,287(51):43071-43082
ADAM17, which is also known as TNFα-converting enzyme, is the major sheddase for the EGF receptor ligands and is considered to be one of the main proteases responsible for the ectodomain shedding of surface proteins. How a membrane-anchored proteinase with an extracellular catalytic domain can be activated by inside-out regulation is not completely understood. We characterized thioredoxin-1 (Trx-1) as a partner of the ADAM17 cytoplasmic domain that could be involved in the regulation of ADAM17 activity. We induced the overexpression of the ADAM17 cytoplasmic domain in HEK293 cells, and ligands able to bind this domain were identified by MS after protein immunoprecipitation. Trx-1 was also validated as a ligand of the ADAM17 cytoplasmic domain and full-length ADAM17 recombinant proteins by immunoblotting, immunolocalization, and solid phase binding assay. In addition, using nuclear magnetic resonance, it was shown in vitro that the titration of the ADAM17 cytoplasmic domain promotes changes in the conformation of Trx-1. The MS analysis of the cross-linked complexes showed cross-linking between the two proteins by lysine residues. To further evaluate the functional role of Trx-1, we used a heparin-binding EGF shedding cell model and observed that the overexpression of Trx-1 in HEK293 cells could decrease the activity of ADAM17, activated by either phorbol 12-myristate 13-acetate or EGF. This study identifies Trx-1 as a novel interaction partner of the ADAM17 cytoplasmic domain and suggests that Trx-1 is a potential candidate that could be involved in ADAM17 activity regulation. 相似文献
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Diane C. Munday Weining Wu Nikki Smith Jenna Fix Sarah Louise Noton Marie Galloux Olivier Touzelet Stuart D. Armstrong Jenna M. Dawson Waleed Aljabr Andrew J. Easton Marie-Anne Rameix-Welti Andressa Peres de Oliveira Fernando M. Simabuco Armando M. Ventura David J. Hughes John N. Barr Rachel Fearns Paul Digard Jean-Fran?ois Eléou?t Julian A. Hiscox 《Journal of virology》2015,89(2):917-930
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Nascimento Filho V. F. Poblete V. H. Parreira P. S. Matsumoto E. Simabuco S. M. Espinoza E. P. Navarro A. A. 《Biological trace element research》1999,(1):423-430
An X-ray tube with a Mo target and Zr filter, operated at 45 kV/20 mA, was used to excite samples (5 ΜL deposited on a quartz
support) and the total reflection angle condition was obtained with a double reflector module built with two 10-cm-long 7-mm-thick
quartz crystals placed 50 Μm apart. A high-resolution spectrometer based on a Si(Li) detector coupled to a multichannel analyzer
was used for X-ray detection and the spectra were interpreted with the AXIL software.
The system was calibrated with standard chemical solutions containing Cr, Fe, Cu, Zn, and Pb, and Y was used as an internal
standard to correct eventual geometric errors and high-voltage instabilities of the X-ray generator. The limits of detection
were 19, 9, 5, and 4 ng/mL for Cr, Fe, Cu, and Zn, respectively, analyzed through characteristicKk
α
X-rays, and 7 ng/mL for Pb, throughLk
α
X-rays, considering 50 ΜL samples deposited and dried on a quartz support, to be excited/ detected for 1000 s. 相似文献
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Morello LG Coltri PP Quaresma AJ Simabuco FM Silva TC Singh G Nickerson JA Oliveira CC Moore MJ Zanchin NI 《PloS one》2011,6(12):e29174
NIP7 is one of the many trans-acting factors required for eukaryotic ribosome biogenesis, which interacts with nascent pre-ribosomal particles and dissociates as they complete maturation and are exported to the cytoplasm. By using conditional knockdown, we have shown previously that yeast Nip7p is required primarily for 60S subunit synthesis while human NIP7 is involved in the biogenesis of 40S subunit. This raised the possibility that human NIP7 interacts with a different set of proteins as compared to the yeast protein. By using the yeast two-hybrid system we identified FTSJ3, a putative ortholog of yeast Spb1p, as a human NIP7-interacting protein. A functional association between NIP7 and FTSJ3 is further supported by colocalization and coimmunoprecipitation analyses. Conditional knockdown revealed that depletion of FTSJ3 affects cell proliferation and causes pre-rRNA processing defects. The major pre-rRNA processing defect involves accumulation of the 34S pre-rRNA encompassing from site A' to site 2b. Accumulation of this pre-rRNA indicates that processing of sites A0, 1 and 2 are slower in cells depleted of FTSJ3 and implicates FTSJ3 in the pathway leading to 18S rRNA maturation as observed previously for NIP7. The results presented in this work indicate a close functional interaction between NIP7 and FTSJ3 during pre-rRNA processing and show that FTSJ3 participates in ribosome synthesis in human cells. 相似文献
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Novel Processed Form of Syndecan-1 Shed from SCC-9 Cells Plays a Role in Cell Migration 总被引:1,自引:0,他引:1
AZ Aragão M Belloni FM Simabuco MR Zanetti S Yokoo RR Domingues R Kawahara BA Pauletti A Gonçalves M Agostini E Graner RD Coletta JW Fox AF Leme 《PloS one》2012,7(8):e43521
The extracellular milieu is comprised in part by products of cellular secretion and cell surface shedding. The presence of such molecules of the sheddome and secretome in the context of the extracellular milieu may have important clinical implications. In cancer they have been hypothesized to play a role in tumor growth and metastasis. The objective of this study was to evaluate whether the sheddome/secretome from two cell lines could be correlated with their potential for tumor development. Two epithelial cell lines, HaCaT and SCC-9, were chosen based on their differing abilities to form tumors in animal models of tumorigenesis. These cell lines when stimulated with phorbol-ester (PMA) showed different characteristics as assessed by cell migration, adhesion and higher gelatinase activity. Proteomic analysis of the media from these treated cells identified interesting, functionally relevant differences in their sheddome/secretome. Among the shed proteins, soluble syndecan-1 was found only in media from stimulated tumorigenic cells (SCC-9) and its fragments were observed in higher amount in the stimulated tumorigenic cells than stimulated non-tumorigenic cells (HaCaT). The increase in soluble syndecan-1 was associated with a decrease in membrane-bound syndecan-1 of SCC-9 cells after PMA stimuli. To support a functional role for soluble syndecan-1 fragments we demonstrated that the synthetic syndecan-1 peptide was able to induce cell migration in both cell lines. Taken together, these results suggested that PMA stimulation alters the sheddome/secretome of the tumorigenic cell line SCC-9 and one such component, the syndecan-1 peptide identified in this study, was revealed to promote migration in these epithelial cell lines. 相似文献