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Miniero DV Cappello AR Curcio R Ludovico A Daddabbo L Stipani I Robinson AJ Kunji ER Palmieri F 《Biochimica et biophysica acta》2011,1807(3):302-310
The mitochondrial oxoglutarate carrier belongs to the mitochondrial carrier family and exchanges oxoglutarate for malate and other dicarboxylates across the mitochondrial inner membrane. Here, single-cysteine mutant carriers were engineered for every residue in the amino- and carboxy-terminus, cytoplasmic loops, and matrix alpha-helices and their transport activity was measured in the presence and absence of sulfhydryl reagents. The analysis of the cytoplasmic side of the oxoglutarate carrier showed that the conserved and symmetric residues of the mitochondrial carrier motif [DE]XX[RK] localized at the C-terminal end of the even-numbered transmembrane alpha-helices are important for the function of the carrier, but the non-conserved cytoplasmic loops and termini are not. On the mitochondrial matrix side of the carrier most residues of the three matrix alpha-helices that are in the interface with the transmembrane alpha-helical bundle are important for function. Among these are the residues of the symmetric [ED]G motif present at the C-terminus of the matrix alpha-helices; the tyrosines of the symmetric YK motif at the N-terminus of the matrix alpha-helices; and the hydrophobic residues M147, I171 and I247. The functional role of these residues was assessed in the structural context of the homology model of OGC. Furthermore, in this study no evidence was found for the presence of a specific homo-dimerisation interface on the surface of the carrier consisting of conserved, asymmetric and transport-critical residues. 相似文献
13.
Marília DV Braga Christian Gautier Marie-France Sagot 《Algorithms for molecular biology : AMB》2009,4(1):16-11
Background
The reversal distance and optimal sequences of reversals to transform a genome into another are useful tools to analyse evolutionary scenarios. However, the number of sequences is huge and some additional criteria should be used to obtain a more accurate analysis. One strategy is searching for sequences that respect constraints, such as the common intervals (clusters of co-localised genes). Another approach is to explore the whole space of sorting sequences, eventually grouping them into classes of equivalence. Recently both strategies started to be put together, to restrain the space to the sequences that respect constraints. In particular an algorithm has been proposed to list classes whose sorting sequences do not break the common intervals detected between the two inital genomes A and B. This approach may reduce the space of sequences and is symmetric (the result of the analysis sorting A into B can be obtained from the analysis sorting B into A). 相似文献14.
Hernández-Quintero JD MH Reyes-Valdés DV Mendoza-Rodríguez M Gómez-Martínez R Rodríguez-Herrera 《Phyton》2015,84(1):107-112
The genus Dasylirion is a group of plants typically present in the Chihuahuan Desert, perennial, with a dioecious sexual behavior and commonly called sotoles. This genus has been little studied from the biological point of view, and the bases of its reproductive response remain unknown. In this work we studied the chromosome number and meiotic response of Dasylirion cedrosanum in the county of Saltillo, Coahuila, located at the North East of Mexico. For the preparation of mitotic chromosomes, we used a technique based on enzymatic treatment with pectolyase and cellulase, as well as staining with acetocarmin dye. For the study of meiosis, male flower buds were collected, fixed and stained for analysis with the same dye. As a result, the gametic (n = x = 19) and somatic chromosome (2n = 38) numbers of D. cedrosanum are reported for the first time, being consistent with previous findings in other Dasylirion species, which points to a constant ploidy level across the genus. Variation was observed in the morphology and size of the somatic chromosomes, with types ranging from submetacentric to subtelocentric, and sizes oscillating in a range of 4.43 µm, with an average total length of 112.38 µm for the diploid chromosome complement. This shows that the chromosome complement of D. cedrosanom would belong to a 3B classification of Stebins, with a medium variation between chromosome lengths and low chromosome asymmetry. This variation indicates the feasibility of constructing a chromosome ideotype for this species. The meiotic chromosome pairing showed a chromosome behavior consistent with a disomic inheritance characteristic of a diploid species, with prevalence of ring and chain bivalents, typically without pairing abnormalities. Bivalent configurations in all cases were symmetrical.The normal and symmetrical meiotic pairing indicates a balanced production of gametes, and suggests the absence of heteromorphic sex determination. 相似文献
15.
Yoshihito Kishita Aleksandra Pajak Nikhita?Ajit Bolar Carlo?M.T. Marobbio Camilla Maffezzini Daniela?V. Miniero Magnus Monné Masakazu Kohda Henrik Stranneheim Kei Murayama Karin Naess Nicole Lesko Helene Bruhn Arnaud Mourier Rolf Wibom Inger Nennesmo Ann Jespers Paul Govaert Akira Ohtake Lut Van?Laer Bart?L. Loeys Christoph Freyer Ferdinando Palmieri Anna Wredenberg Yasushi Okazaki Anna Wedell 《American journal of human genetics》2015,97(5):761-768
S-adenosylmethionine (SAM) is the predominant methyl group donor and has a large spectrum of target substrates. As such, it is essential for nearly all biological methylation reactions. SAM is synthesized by methionine adenosyltransferase from methionine and ATP in the cytoplasm and subsequently distributed throughout the different cellular compartments, including mitochondria, where methylation is mostly required for nucleic-acid modifications and respiratory-chain function. We report a syndrome in three families affected by reduced intra-mitochondrial methylation caused by recessive mutations in the gene encoding the only known mitochondrial SAM transporter, SLC25A26. Clinical findings ranged from neonatal mortality resulting from respiratory insufficiency and hydrops to childhood acute episodes of cardiopulmonary failure and slowly progressive muscle weakness. We show that SLC25A26 mutations cause various mitochondrial defects, including those affecting RNA stability, protein modification, mitochondrial translation, and the biosynthesis of CoQ10 and lipoic acid. 相似文献
16.
Magnus Monné Lucia Daddabbo Lorena Carla Giannossa Maria Cristina Nicolardi Luigi Palmieri Daniela Valeria Miniero Annarosa Mangone Ferdinando Palmieri 《Journal of bioenergetics and biomembranes》2017,49(5):369-380
The ATP-Mg/phosphate carriers (APCs) modulate the intramitochondrial adenine nucleotide pool size. In this study the concentration-dependent effects of Mg2+ and other divalent cations (Me2+) on the transport of [3H]ATP in liposomes reconstituted with purified human and Arabidopsis APCs (hAPCs and AtAPCs, respectively, including some lacking their N-terminal domains) have been investigated. The transport of Me2+ mediated by these proteins was also measured. In the presence of a low external concentration of [3H]ATP (12 μM) and increasing concentrations of Me2+, Mg2+ stimulated the activity (measured as initial transport rate of [3H]ATP) of hAPCs and decreased that of AtAPCs; Fe2+ and Zn2+ stimulated markedly hAPCs and moderately AtAPCs; Ca2+ and Mn2+ markedly AtAPCs and moderately hAPCs; and Cu2+ decreased the activity of both hAPCs and AtAPCs. All the Me2+-dependent effects correlated well with the amount of ATP-Me complex present. The transport of [14C]AMP, which has a much lower ability of complexation than ATP, was not affected by the presence of the Me2+ tested, except Cu2+. Furthermore, the transport of [3H]ATP catalyzed by the ATP/ADP carrier, which is known to transport only free ATP and ADP, was inhibited by all the Me2+ tested in an inverse relationship with the formation of the ATP-Me complex. Finally, direct measurements of Mg2+, Mn2+, Fe2+, Zn2+ and Cu2+ showed that they are cotransported with ATP by both hAPCs and AtAPCs. It is likely that in vivo APCs transport free ATP and ATP-Mg complex to different degrees, and probably trace amounts of other Me2+ in complex with ATP. 相似文献
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Monné M Miniero DV Daddabbo L Robinson AJ Kunji ER Palmieri F 《The Journal of biological chemistry》2012,287(11):7925-7934
Mitochondrial carriers are a large family of proteins that transport specific metabolites across the inner mitochondrial membrane. Sequence and structure analysis has indicated that these transporters have substrate binding sites in a similar location of the central cavity consisting of three major contact points. Here we have characterized mutations of the proposed substrate binding site in the human ornithine carriers ORC1 and ORC2 by carrying out transport assays with a set of different substrates. The different substrate specificities of the two isoforms, which share 87% identical amino acids, were essentially swapped by exchanging a single residue located at position 179 that is arginine in ORC1 and glutamine in ORC2. Altogether the substrate specificity changes demonstrate that Arg-179 and Glu-180 of contact point II bind the C(α) carboxylate and amino group of the substrates, respectively. Residue Glu-77 of contact point I most likely interacts with the terminal amino group of the substrate side chain. Furthermore, it is likely that all three contact points are involved in the substrate-induced conformational changes required for substrate translocation because Arg-179 is probably connected with Arg-275 of contact point III through Trp-224 by cation-π interactions. Mutations at position 179 also affected the turnover number of the ornithine carrier severely, implying that substrate binding to residue 179 is a rate-limiting step of the catalytic transport cycle. Given that Arg-179 is located in the vicinity of the matrix gate, it is concluded that it is a key residue in the opening of the carrier to the matrix side. 相似文献
19.
The timing of alpha-gustducin expression during cell renewal in rat vallate taste buds 总被引:2,自引:2,他引:0
The G protein subunit alpha-gustducin is expressed in a subset of light
(Type II) but not in dark (Type I) cells in rat vallate taste buds. The
thymidine analogue 5-bromo-2'-deoxyuridine (BrdU) is incorporated into DNA
during the S-phase of the cell cycle and can be used to determine the time
of origin of a cell. In this study, 31 rats were injected with BrdU (50
mg/kg i.p.) and perfused at various times, from 2.5 to 10.5 days, following
BrdU administration. Vallate papillae were embedded in polyester wax, cut
into 4 microm transverse sections, and characterized with antibodies to
BrdU and alpha-gustducin. Sections were processed for indirect
immunofluorescence or with an immunoperoxidase procedure. From
immunoperoxidase material on 21 rats, counts of alpha-gustducin- and
BrdU-labeled cells were obtained from 300-800 taste bud profiles at each
survival time; a total of 4122 taste bud profiles were examined. Cells with
nuclei immunoreactive for BrdU occurred within the taste buds at 2.5 days
and double-labeled cells were clearly evident at 3.5 days; a small number
of double-labeled cells were seen as early as 2.5 days. Double-labeled
cells reached a peak at 6.5 days and did not decline significantly by 10.5
days. Cells labeled for BrdU but not alpha-gustducin peaked at 5.5 days and
showed a significant decline by 8.5 days. These latter cells included light
cells not expressing alpha- gustducin and dark cells, which have previously
been shown to have a shorter life span than light cells. These data suggest
that expression of alpha-gustducin appears very early in a cell's life span
and that these cells are longer lived than many of the cells that do not
express this G protein.
相似文献
20.
Tonic GABAergic inhibition of taste-responsive neurons in the nucleus of the solitary tract 总被引:4,自引:1,他引:3
The effects of gamma-aminobutyric acid (GABA) and the GABAA receptor
antagonist bicuculline methiodide (BICM) on the activity of taste-
responsive neurons in the nucleus of the solitary tract (NST) were examined
electrophysiologically in urethane-anesthetized hamsters. Single neurons in
the NST were recorded extracellularly and drugs (21 nl) were microinjected
into the vicinity of the cell via a multibarrel pipette. The response of
each cell was recorded to lingual stimulation with 0.032 M NaCl, 0.032 M
sucrose, 0.0032 M citric acid and 0.032 M quinine hydrochloride (QHCl).
Forty-six neurons were tested for the effects of GABA; the activity of 29
cells (63%) was inhibited by 5 mM GABA. Whether activity was elicited in
these cells by repetitive anodal current stimulation (25 microA, 0.5 s, 0.1
Hz) of the tongue (n = 13 cells) or the cells were spontaneously active (n
= 13 cells), GABA produced a dose-dependent (1, 2 and 5 mM) decrement in
activity. Forty- seven NST neurons were tested for the effects of BICM on
their responses to chemical stimulation of the tongue; the responses of 28
cells (60%) were enhanced by 10 mM BICM. The gustatory responses of 26 of
these cells were tested with three concentrations (0.2, 2 and 10 mM) of
BICM, which produced a dose-dependent increase in both spontaneous activity
and taste-evoked responses. Nine of these neurons were sucrose- best, seven
were NaCl-best, eight were acid-best and two responded best to QHCl. The
responses to all four tastants were enhanced, with no difference among
neuron types. For 18 cells that were tested with two or more gustatory
stimuli, BICM increased their breadth of responsiveness to their two most
effective stimuli. These data show that approximately 60% of the
taste-responsive neurons in the rostral NST are inhibited by GABA and/or
subject to a tonic inhibitory influence, which is mediated by GABAA
receptors. The modulation of these cells by GABA provides a mechanism by
which the breadth of tuning of the cell can be sharpened. Modulation of
gustatory activity following a number of physiological changes could be
mediated by such a GABAergic circuit.
相似文献