Investigation of the spatial distribution of glutathione redox-balance in live cells by using Fluorescence Ratio Imaging Microscopy

Maulucci, Giuseppe; Pani, Giovambattista; Labate, Valentina; Mele, M.C.; Panieri, Emiliano; Papi, Massimiliano; Arcòvito, Giuseppe; Galeotti, T.; Spirito, Marco De

doi:10.1016/j.bios.2009.07.038

Cited by 26 publications

(14 citation statements)

References 17 publications

(14 reference statements)

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“…Two-photon infrared excitation techniques have been successfully applied to detect Laurdan emission [22]. By using this probe, coexisting lipid domains were characterized on the basis of their distinctive fluorescence spectra and dual-wavelength ratio measurements [23][24][25], which map changes in the structure of PM [19,22,[26][27][28].…”

Section: Introductionmentioning

confidence: 99%

Fatty acid-related modulations of membrane fluidity in cells: detection and implications

Maulucci

Cohen

Daniel

et al. 2016

Free Radical Research

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129

101

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Metabolic homeostasis of fatty acids is complex and well-regulated in all organisms. The biosynthesis of saturated fatty acids (SFA) in mammals provides substrates for b-oxidation and ATP production. Monounsaturated fatty acids (MUFA) are products of desaturases that introduce a methylene group in cis geometry in SFA. Polyunsaturated fatty acids (n-6 and n-3 PUFA) are products of elongation and desaturation of the essential linoleic acid and a-linolenic acid, respectively. The liver processes dietary fatty acids and exports them in lipoproteins for distribution and storage in peripheral tissues. The three types of fatty acids are integrated in membrane phospholipids and determine their biophysical properties and functions. This study was aimed at investigating effects of fatty acids on membrane biophysical properties under varying nutritional and pathological conditions, by integrating lipidomic analysis of membrane phospholipids with functional two-photon microscopy (fTPM) of cellular membranes. This approach was applied to two case studies: first, pancreatic beta-cells, to investigate hormetic and detrimental effects of lipids. Second, red blood cells extracted from a genetic mouse model defective in lipoproteins, to understand the role of lipids in hepatic diseases and metabolic syndrome and their effect on circulating cells.ARTICLE HISTORY

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Section: Introductionmentioning

confidence: 99%

Fatty acid-related modulations of membrane fluidity in cells: detection and implications

Maulucci

Cohen

Daniel

et al. 2016

Free Radical Research

Self Cite

129

101

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“…As fundamental cellular metabolic processes and efficient redox signaling can be guaranteed if redox homeostasis is optimally maintained (Meyer, 2008), there must exist a delicate mechanism that regulates redox homeostasis in plant cells. Given that: (1) ROS metabolism is known to be a major factor in the control of cellular redox homeostasis; (2) excess ROS is produced not only in response to environmental stresses, but also during certain developmental processes under normal conditions; and (3) ROS production is compartmentalized, with some plastids, including chloroplasts, being the major ROS production sites (Hansen et al, 2006;Maulucci et al, 2009;Ushio-Fukai, 2009), the transfer of ZmRIP1 from the chloroplast to the nucleus in response to H 2 O 2 appears to suggest a feedback mechanism for regular regulation of redox homeostasis in maize cells.…”

Section: Discussionmentioning

confidence: 99%

Purification and Characterization of ZmRIP1, a Novel Reductant-Inhibited Protein Tyrosine Phosphatase from Maize

Zhao²,

Liang³

et al. 2012

Plant Physiology

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Protein tyrosine phosphatases (PTPases) have long been thought to be activated by reductants and deactivated by oxidants, owing to the presence of a crucial sulfhydryl group in their catalytic centers. In this article, we report the purification and characterization of Reductant-Inhibited PTPase1 (ZmRIP1) from maize (Zea mays) coleoptiles, and show that this PTPase has a unique mode of redox regulation and signaling. Surprisingly, ZmRIP1 was found to be deactivated by a reductant. A cysteine (Cys) residue (Cys-181) near the active center was found to regulate this unique mode of redox regulation, as mutation of Cys-181 to arginine-181 allowed ZmRIP1 to be activated by a reductant. In response to oxidant treatment, ZmRIP1 was translocated from the chloroplast to the nucleus. Expression of ZmRIP1 in Arabidopsis (Arabidopsis thaliana) plants and maize protoplasts altered the expression of genes encoding enzymes involved in antioxidant catabolism, such as At1g02950, which encodes a glutathione transferase. Thus, the novel PTPase identified in this study is predicted to function in redox signaling in maize.

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“…Imaging was performed at room temperature. Image processing was performed with ImageJ software; image background values (defined as intensities below 7% of the maximum intensity) were set to zero and colored in black [33]. Intensity profiles were measured on bacteria entire length with ‘line profile’ tool [34].…”

Section: Methodsmentioning

confidence: 99%

Impact of Protein Domains on PE_PGRS30 Polar Localization in Mycobacteria

et al. 2014

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PE_PGRS proteins are unique to the Mycobacterium tuberculosis complex and a number of other pathogenic mycobacteria. PE_PGRS30, which is required for the full virulence of M. tuberculosis (Mtb), has three main domains, i.e. an N-terminal PE domain, repetitive PGRS domain and the unique C-terminal domain. To investigate the role of these domains, we expressed a GFP-tagged PE_PGRS30 protein and a series of its functional deletion mutants in different mycobacterial species (Mtb, Mycobacterium bovis BCG and Mycobacterium smegmatis) and analysed protein localization by confocal microscopy. We show that PE_PGRS30 localizes at the mycobacterial cell poles in Mtb and M. bovis BCG but not in M. smegmatis and that the PGRS domain of the protein strongly contributes to protein cellular localization in Mtb. Immunofluorescence studies further showed that the unique C-terminal domain of PE_PGRS30 is not available on the surface, except when the PGRS domain is missing. Immunoblot demonstrated that the PGRS domain is required to maintain the protein strongly associated with the non-soluble cellular fraction. These results suggest that the repetitive GGA-GGN repeats of the PGRS domain contain specific sequences that contribute to protein cellular localization and that polar localization might be a key step in the PE_PGRS30-dependent virulence mechanism.

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Investigation of the spatial distribution of glutathione redox-balance in live cells by using Fluorescence Ratio Imaging Microscopy

Cited by 26 publications

References 17 publications

Fatty acid-related modulations of membrane fluidity in cells: detection and implications

Fatty acid-related modulations of membrane fluidity in cells: detection and implications

Purification and Characterization of ZmRIP1, a Novel Reductant-Inhibited Protein Tyrosine Phosphatase from Maize

Impact of Protein Domains on PE_PGRS30 Polar Localization in Mycobacteria

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