Alexei Grichine
French Institute of Health and Medical Research
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Publication
Featured researches published by Alexei Grichine.
Journal of the American Chemical Society | 2008
Alexandre Picot; Anthony D'Aléo; Patrice L. Baldeck; Alexei Grichine; Alain Duperray; Chantal Andraud; Olivier Maury
A new europium complex presenting good solubility and stability in water, intense emission in the red (616 nm), long luminescence lifetime, and significant two-photon absorption cross-section in the biological window has been designed and successfully used for two-photon scanning microscopy bioimaging experiments on fixed cancer cells.
Molecular Biology of the Cell | 2009
Anton Sandqvist; Johanna K. Björk; Malin Åkerfelt; Zhanna Chitikova; Alexei Grichine; Claire Vourc'h; Caroline Jolly; Tiina A. Salminen; Yvonne Nymalm; Lea Sistonen
Organisms respond to circumstances threatening the cellular protein homeostasis by activation of heat-shock transcription factors (HSFs), which play important roles in stress resistance, development, and longevity. Of the four HSFs in vertebrates (HSF1-4), HSF1 is activated by stress, whereas HSF2 lacks intrinsic stress responsiveness. The mechanism by which HSF2 is recruited to stress-inducible promoters and how HSF2 is activated is not known. However, changes in the HSF2 expression occur, coinciding with the functions of HSF2 in development. Here, we demonstrate that HSF1 and HSF2 form heterotrimers when bound to satellite III DNA in nuclear stress bodies, subnuclear structures in which HSF1 induces transcription. By depleting HSF2, we show that HSF1-HSF2 heterotrimerization is a mechanism regulating transcription. Upon stress, HSF2 DNA binding is HSF1 dependent. Intriguingly, when the elevated expression of HSF2 during development is mimicked, HSF2 binds to DNA and becomes transcriptionally competent. HSF2 activation leads to activation of also HSF1, revealing a functional interdependency that is mediated through the conserved trimerization domains of these factors. We propose that heterotrimerization of HSF1 and HSF2 integrates transcriptional activation in response to distinct stress and developmental stimuli.
Journal of Cell Biology | 2008
Angélique Millon-Frémillon; Daniel Bouvard; Alexei Grichine; Sandra Manet-Dupé; Marc R. Block; Corinne Albiges-Rizo
Cell migration is an integrated process requiring the continuous coordinated assembly and disassembly of adhesion structures. How cells orchestrate adhesion turnover is only partially understood. We provide evidence for a novel mechanistic insight into focal adhesion (FA) dynamics by demonstrating that integrin cytoplasmic domain–associated protein 1 (ICAP-1) slows down FA assembly. Live cell imaging, which was performed in both Icap-1–deficient mouse embryonic fibroblasts and cells expressing active β1 integrin, shows that the integrin high affinity state favored by talin is antagonistically controlled by ICAP-1. This affinity switch results in modulation in the speed of FA assembly and, consequently, of cell spreading and migration. Unexpectedly, the ICAP-1–dependent decrease in integrin affinity allows cell sensing of matrix surface density, suggesting that integrin conformational changes are important in mechanotransduction. Our results clarify the function of ICAP-1 in cell adhesion and highlight the central role it plays in the cells integrated response to the extracellular microenvironment.
Angewandte Chemie | 2012
Anthony D'Aléo; Adrien Bourdolle; Sophie Brustlein; Teddy Fauquier; Alexei Grichine; Alain Duperray; Patrice L. Baldeck; Chantal Andraud; Sophie Brasselet; Olivier Maury
HAL is a multi-disciplinary open access archive for the deposit and dissemination of scientific research documents, whether they are published or not. The documents may come from teaching and research institutions in France or abroad, or from public or private research centers. L’archive ouverte pluridisciplinaire HAL, est destinée au dépôt et à la diffusion de documents scientifiques de niveau recherche, publiés ou non, émanant des établissements d’enseignement et de recherche français ou étrangers, des laboratoires publics ou privés. Ytterbium-based bioprobes for near-infrared two-photon scanning laser microscopy imaging. Anthony D’Aléo, Adrien Bourdolle, Sophie Brustlein, Teddy Fauquier, Alexei Grichine, Alain Duperray, P. L. Baldeck, Chantal Andraud, Sophie Brasselet, Olivier Maury
Biochimica et Biophysica Acta | 2012
Rita Guzun; Marcela Gonzalez-Granillo; Minna Karu-Varikmaa; Alexei Grichine; Yves Usson; Tuuli Kaambre; Karen Guerrero-Roesch; Andrey V. Kuznetsov; Uwe Schlattner; Valdur Saks
This review describes the recent experimental data on the importance of the VDAC-cytoskeleton interactions in determining the mechanisms of energy and metabolite transfer between mitochondria and cytoplasm in cardiac cells. In the intermembrane space mitochondrial creatine kinase connects VDAC with adenine nucleotide translocase and ATP synthase complex, on the cytoplasmic side VDAC is linked to cytoskeletal proteins. Applying immunofluorescent imaging and Western blot analysis we have shown that β2-tubulin coexpressed with mitochondria is highly important for cardiac muscle cells mitochondrial metabolism. Since it has been shown by Rostovtseva et al. that αβ-heterodimer of tubulin binds to VDAC and decreases its permeability, we suppose that the β-tubulin subunit is bound on the cytoplasmic side and α-tubulin C-terminal tail is inserted into VDAC. Other cytoskeletal proteins, such as plectin and desmin may be involved in this process. The result of VDAC-cytoskeletal interactions is selective restriction of the channel permeability for adenine nucleotides but not for creatine or phosphocreatine that favors energy transfer via the phosphocreatine pathway. In some types of cancer cells these interactions are altered favoring the hexokinase binding and thus explaining the Warburg effect of increased glycolytic lactate production in these cells. This article is part of a Special Issue entitled: VDAC structure, function, and regulation of mitochondrial metabolism.
Biochimie | 2011
Leilei Zhang; Minh Vu Chuong Nguyen; Bernard Lardy; Algirdas J. Jesaitis; Alexei Grichine; Francis Rousset; Monique Talbot; Marie-Hélène Paclet; GuanXiang Qian; Françoise Morel
Nox4, a member of Nox family of NADPH oxidase expressed in nonphagocytic cells, is a major source of reactive oxygen species in many cell types. But understanding of the role of Nox4 in the production of ROS and of regulation mechanism of oxidase activity is largely unknown. This study reports for the first time the generation and characterization of 5 mAbs against a recombinant Nox4 protein (AA: 206-578). Among 5 novel mAbs, 3 mAbs (8E9, 5F9, 6B11) specifically recognized Nox4 protein in HEK293 transfected cells or human kidney cortex by western blot analysis; mAb 8E9 reacted with intact tet-induced T-REx™ Nox4 cells in FACS studies. The other 2 mAbs 10B4 and 7C9 were shown to have a very weak reactivity after purification. Immunofluorescence confocal microscopy showed that Nox4 localized not only in the perinuclear and endoplasmic reticulum regions but also at the plasma membrane of the cells which was further confirmed by TIRF-microscopy. Epitope determination showed that mAb 8E9 recognizes a region on the last extracellular loop of Nox4, while mAbs 6B11 and 5F9 are directed to its cytosolic tail. Contrary to mAb 6B11, mAb 5F9 failed to detect Nox4 at the plasma membrane. Cell-free oxidase assays demonstrated a moderate but significant inhibition of constitutive Nox4 activity by mAbs 5F9 and 6B11. In conclusion, 5 mAbs raised against Nox4 were generated for the first time. 3 of them will provide powerful tools for a structure/function relationship of Nox4 and for physiopathological investigations in humans.
Chemical Science | 2014
Alexei Grichine; Alexandre Haefele; Simon Pascal; Alain Duperray; Richard Michel; Chantal Andraud; Olivier Maury
The long luminescence lifetime of lanthanide based bioprobes is a great advantage for their specific detection in autofluorescent or labelled cells and tissues. It is also a valuable tool for sensing the physicochemical microenvironment and molecular interactions by Forster resonance energy transfer (FRET). However, standard confocal and multiphoton laser scanning microscopes are not adapted for imaging with such temporal resolution, because the typical pixel dwell time is too short compared to the luminescence lifetime. We show that the rapid sampling rate and laser control of a usual confocal microscope can instead be used for precise measurement of long lifetime decays (μs to ms range). Furthermore, both raster- and line-scanning microscopes can specifically detect long luminescence signals in the time-gated mode by shifting the pinhole or the confocal slit in the lagging direction. We characterized the subcellular localization and accurately measured the millisecond luminescence lifetimes of the benchmark two-photon europium probe [Na]3[EuL1G3], and specifically imaged this label in the presence of short-lived fluorescent species. Fine variations of the luminescence lifetime of this lanthanide complex were revealed and mapped in cells in the presence of a FRET acceptor, allowing quantification of the FRET efficiency independently of donor concentration. These results demonstrate a high and yet unexploited potential of quantitative confocal and multiphoton microscopy for time-gated and lifetime imaging of lanthanide-based biological sensors.
Molecular Imaging | 2007
Zhao-Hui Jin; Jesus Razkin; Véronique Josserand; Didier Boturyn; Alexei Grichine; Isabelle Texier; Marie-Christine Favrot; Pascal Dumy; Jean-Luc Coll
We reported that regioselectively addressable functionalized template (RAFT)-c(-RGDfK-)4 presenting four cyclic (Arg-Gly-Asp) (cRGD) peptides targets integrin aVb3 with an improved specificity compared with monomeric cRGD. In this study, we improved this vector by creating a “stealth” molecule in which a fluorescence quencher (Q) is linked to Cy5 via a disulfide bond (-SS-). RAFT-c(-RGDfK-)4-Cy5-SS-Q fluorescence is quenched unless activated by reduction during internalization. RAFT-c(-RGDfK-)4-Cy5-SS-Q fluorescence was negligible when compared with the control but totally recovered after cleavage of the disulfide bridge. Confocal microscopy showed that only the intracellular Cy5 signal could be detected using RAFT-c(-RGDfK-)4-Cy5-SS-Q, confirming that uncleaved extracellular molecules are not visible. Whole-body imaging of mice bearing subcutaneous tumors injected intravenously with RAFT-c(-RGDfK-)4-Cy5-SS-Q showed a very significant enhancement of the fluorescent contrast in tumors compared with the unquenched molecule. Histology of the tumor confirmed the intracellular accumulation of Cy5. These results demonstrate that the presence of a labile disulfide bridge between the targeting vector and a drug mimetic is an efficient way to deliver a dye, or a drug, intracellularly. In addition, this quenched RAFT-c(-RGDfK-)4-Cy5-SS-Q probe is a very powerful vector for imaging tumor masses and investigating in vivo RGD-mediated internalization.
Journal of Biotechnology | 2010
Virgile Adam; Hideaki Mizuno; Alexei Grichine; Jun-ichi Hotta; Yutaka Yamagata; Benjamien Moeyaert; G. Ulrich Nienhaus; Atsushi Miyawaki; Dominique Bourgeois; Johan Hofkens
The recent discovery of photoconvertible and photoswitchable fluorescent proteins (PCFPs and RSFPs, respectively) that can undergo photoinduced changes of their absorption/emission spectra opened new research possibilities in subdiffraction microscopy and optical data storage. Here we demonstrate the proof-of-principle for read only and rewritable data storage both in 2D and 3D, using PCFPs and RSFPs. The irreversible burning of information was achieved by photoconverting from green to red defined areas in a layer of the PCFP Kaede. Data were also written and erased several times in layers of the photochromic fluorescent protein Dronpa. Using IrisFP, which combines the properties of PCFPs and RSFPs, we performed the first encoding of data in four colours using only one type of fluorescent protein. Finally, three-dimensional optical data storage was demonstrated using three mutants of EosFP (d1EosFP, mEosFP and IrisFP) in their crystalline form. Two-photon excitation allowed the precise addressing of regions of interest (ROIs) within the three-dimensional crystalline matrix without excitation of out-of-focus optical planes. Hence, this contribution highlights several data storage schemes based on the remarkable properties of PCFPs/RSFPs.
Journal of Cell Biology | 2014
Boubou Diagouraga; Alexei Grichine; Arnold Fertin; Jin Wang; Saadi Khochbin; Karin Sadoul
During platelet activation, motor protein-induced coiling of the microtubule-based marginal band leads to the cells’ characteristic spherical shape, whereas actomyosin-mediated compression of the coil results in new microtubule polymerization in a smaller ring.