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71.
72.
Dominique Rumeau Stephan Cuiné Laurent Fina Nathalie Gault Michel Nicole Gilles Peltier 《Planta》1996,199(1):79-88
The intracellular compartmentation of carbonic anhydrase (CA; EC 4.2.1.1), an enzyme that catalyses the reversible hydration of CO2 to bicarbonate, has been investigated in potato (Solanum tuberosum L.) leaves. Although enzyme activity was mainly located in chloroplasts (87% of total cellular activity), significant activity (13%) was also found in the cytosol. The corresponding CA isoforms were purified either from chloroplasts or crude leaf extracts, respectively. The cytosolic isoenzyme has a molecular mass of 255 000 and is composed of eight identical subunits with an estimated M
r of 30000. The chloroplastic isoenzyme (M
r 220000) is also an octamer composed of two different subunits with M
r estimated at 27 000 and 27 500, respectively. The N-terminal amino acid sequences of both chloroplastic CA subunits demonstrated that they were identical except that the M
r-27 000 subunit was three amino acids shorter than that of the M
r-27 500 subunit. Cytosolic and chloroplastic CA isoenzymes were found to be similarly inhibited by monovalent anions (Cl–, I–, N
3
-
and NO
3
-
) and by sulfonamides (ethoxyzolamide and acetozolamide). Both CA isoforms were found to be dependent on a reducing agent such as cysteine or dithiothreitol in order to retain the catalytic activity, but 2-mercaptoethanol was found to be a potent inhibitor. A polyclonal antibody directed against a synthetic peptide corresponding to the N-terminal amino acid sequence of the chloroplastic CA monomers also recognized the cytosolic CA isoform. This antibody was used for immunocytolocalization experiments which confirmed the intracellular compartmentation of CA: within chloroplasts, CA is restricted to the stroma and appears randomly distributed in the cytosol.Abbreviations BSA
bovine serum albumin
- CA
carbonic anhydrase
- PMSF
phenylmethylsulphonyl fluoride
- BAM
benzamidine
- DTT
dithiothreitol
- 2-ME
2-mercaptoethanol
- PVDF
polyvinylidene difluoride
The authors thanks P. Carrier and Dr. B. Dimon for technical assistance with the mass-spectrometry measurements. 相似文献
73.
Mapping of a Gene for Long QT Syndrome to Chromosome 4q25-27 总被引:21,自引:0,他引:21
Jean-Jacques Schott Flavien Charpentier Sophie Peltier Patrick Foley Emmanuel Drouin Jean-Brieuc Bouhour Patricia Donnelly Gilles Vergnaud Lucien Bachner Jean-Paul Moisan Herv Le Marec Olivier Pascal 《American journal of human genetics》1995,57(5):1114-1122
Long QT syndrome (LQTS) is a heterogeneous inherited disorder causing syncope and sudden death from ventricular arrhythmias. A first locus for this disorder was mapped to chromosome 11p15.5. However, locus heterogeneity has been demonstrated in several families, and two other loci have recently been located on chromosomes 7q35-36 and 3p21-24. We used linkage analysis to map the locus in a 65-member family in which LQTS was associated with more marked sinus bradycardia than usual, leading to sinus node dysfunction. Linkage to chromosome 11p15.5, 7q35-36, or 3p21-24 was excluded. Positive linkage was obtained for markers located on chromosome 4q25-27. A maximal LOD score of 7.05 was found for marker D4S402. The identification of a fourth locus for LQTS confirms its genetic heterogeneity. Locus 4q25-27 is associated with a peculiar phenotype within the LQTS entity. 相似文献
74.
Josiane Girardie Adrien Girardie Alain Van Dorsselaer Odile Sorokine Serge Geoffre Michel Hospital Gilles Precigoux 《Journal of peptide science》1995,1(5):311-318
An original insect neurohormone of 65 residues was synthesized by the solid-phase methodology using t-Boc strategy and Boc-Val-PAM–resin. The purification, conducted by several steps of liquid chromatography having mass, polarity or charge as separative criteria, yielded the product with the correct molecular weight of 6922 Da determined by mass spectrometry. The synthetic peptide had both the same affinity for the antinative neurohormone serum and the same biological activity as the native neurohormone. 相似文献
75.
Madhulika B. Gupta Maxim D. Seferovic Suya Liu Robert J. Gratton Amanda Doherty-Kirby Gilles A. Lajoie Victor K. M. Han 《Clinical proteomics》2006,2(3-4):169-184
Fetal growth restriction (FGR) affects 3–5% of pregnancies and is associated with increased perinatal morbidity and mortality.
Currently, there is no reliable biochemical test to differentiate a pathological FGR from a nonpathological one. The objective
of this study was to screen whole maternal plasma to identify differentially expressed relatively abundant proteins associated
with FGR. We analyzed maternal plasma from FGR (n=28) and healthy (n=22) pregnancies using two-dimensional gel electrophoresis (2D-GE) followed by software image analysis. Three spots with molecular
weight (Mr) 18 kDa corresponding to haptoglobin (hp) α2, as identified by LC-MS/MS and immunoblotting, showed differential expression
patterns in FGR. The distribution of hp α2 variants in maternal plasma samples showed the hp α2 variant 1 was low in 72% of
FGR, medium in 16%, whereas high in 12%. In comparison, hp α2 variant 1 was high in (41%) of controls, medium in 41%, and
low in 18% of cases. Based on the software image analysis, the mean spot volume for hp α2 variant 1 was 0.12 (SD=0.18) for
FGR compared to 0.26 (SD=0.19) for control (p=0.006). Given that hp turnover is indicative of its maturation process and is traceable in plasma by its dominant/suppressed
variants, we propose that hp α2 is an important potential target for evaluation of its clinical and pathophysiological role
and as a diagnostic biomarker in FGR. 相似文献
76.
Josette Garnier Aurélie Cébron Gaëlle Tallec Gilles Billen Mathieu Sebilo Anun Martinez 《Biogeochemistry》2006,77(3):305-326
The Seine River estuary (France) is the receptacle of a drainage basin characterised by high population density, heavy industrial
activity and intensive agriculture. Whereas nitrate concentrations are high due to diffuse sources in the upstream drainage
basin, ammonium mainly originates from the effluents of the Achères wastewater treatment plant (WWTP) downstream from Paris
and its suburbs (6.5 million equivalent-inhabitants). Ammonium is mostly nitrified in the tidal freshwater estuary and nitrification
causes a strong summer oxygen deficit. Average longitudinal summer profiles of oxygen and nitrogen concentrations for two
periods, between 1993–1997 and 1998–2003 in dry hydrological conditions (excluding the wet years 2000 and 2001) clearly reflect
the changes due to the improved treatment of wastewater from Paris and its suburbs. On the basis of daily water flux data
and twice monthly nitrogen measurements at the boundaries of the upstream freshwater estuarine section (108 km), we calculated
nitrification and denitrification fluxes, whose annual averages were 43 and 71 × 103 kg N d−1 respectively from 1993 to 2003, with summer values (July–September) representing 73 and 57% of the annual fluxes, respectively.
The degree of denitrification in the upper estuary appears to be closely related to the nitrification, itself more loosely
related to the amount of reduced nitrogen (Kjeldahl) brought by the treated effluents from the Achères WWTP. We estimated
the total N2O emissions to about 40 kg N d−1 (25–60 kg N d−1) in the same sector. 相似文献
77.
78.
Colacino E Sindona G Gosselin G Mathé C 《Nucleosides, nucleotides & nucleic acids》2003,22(11):2013-2026
In this article, we describe the synthesis of 5-nitro-1-(2-deoxy-alpha-D-erythro-pentofuranosyl)cytosine (4alpha), 5-nitro-1-(2-deoxy-beta-D-erythro-pentofuranosyl)cytosine (4beta), 5-amino-1-(2-deoxy-alpha-D-erythro-pentofuranosyl)cytosine (5alpha), 5-nitro-1-(2-deoxy-beta-D-erythro-pentofuranosyl)cytosine (5beta), 5-nitro-1-(2,3-dideoxy-beta-D-ribofuranosyl)uracil (6beta), 5-amino-1-(2,3-dideoxy-alpha,beta-D-ribofuranosyl)uracil (7), 5-nitro-1-(2,3-dideoxy-alpha,beta-D-ribofuranosyl)cytosine (8) and 5-amino-1-(2,3-dideoxy-beta-D-ribofuranosyl)cytosine (9beta). The prepared compounds were tested for their activity against HIV and HBV viruses, but they did not show significant activity. 相似文献
79.
Noens EE Mersinias V Willemse J Traag BA Laing E Chater KF Smith CP Koerten HK van Wezel GP 《Molecular microbiology》2007,64(5):1244-1259
80.
Godat E Hervé-Grvépinet V Veillard F Lecaille F Belghazi M Brömme D Lalmanach G 《Biological chemistry》2008,389(8):1123-1126
Although cysteine cathepsins, including cathepsin K, are sensitive to oxidation, proteolytically active forms are found at inflammatory sites. Regulation of cathepsin K activity was analyzed in the presence of H2O2 to gain an insight into these puzzling observations. H2O2 impaired processing of procathepsin K and inactivated its mature form in a time- and dose-dependent mode. However, as a result of the formation of a sulfenic acid, as confirmed by trapping in the presence of 7-chloro-4-nitrobenzo-2-oxa-1,3-diazol, approximately one-third of its initial activity was restored by dithiothreitol. This incomplete inactivation may partially explain why active cysteine cathepsins are still found during acute lung inflammation. 相似文献