RESUMO
We describe the case of a 22 yr old male patient with cystic fibrosis, who, after long-term antibiotic treatment of pulmonary infection, developed a haemorrhagic diathesis with severe bleeding from the mucus membrane of the mouth, and haematuria. Rapid recovery was observed after infusion of vitamin K. During 8 months of follow-up, no evidence of recurrence of the clotting disturbances and anaemia were noted. The combination of impaired absorption of vitamin K due to underlying disease with the antibiotic-induced suppression of vitamin K synthesis by intestinal bacteria could be a possible explanation for this disorder.
Assuntos
Cefuroxima/efeitos adversos , Cefalosporinas/efeitos adversos , Fibrose Cística/complicações , Gentamicinas/efeitos adversos , Transtornos Hemorrágicos/induzido quimicamente , Netilmicina/efeitos adversos , Pneumonia/tratamento farmacológico , Adulto , Cefuroxima/uso terapêutico , Cefalosporinas/uso terapêutico , Fibrose Cística/tratamento farmacológico , Seguimentos , Gentamicinas/uso terapêutico , Transtornos Hemorrágicos/tratamento farmacológico , Transtornos Hemorrágicos/fisiopatologia , Humanos , Masculino , Netilmicina/uso terapêutico , Pneumonia/complicações , Pneumonia/microbiologia , Fatores de Tempo , Vitamina K/uso terapêuticoRESUMO
This study examined, in the largest sample of major depressives reported so far, platelet serotonergic parameters (5-HT uptake, [3H]paroxetine binding and 5-HT2A receptors measured by [3H]LSD binding) in 60 antidepressant-free depressed patients and 40 age- and gender-matched control subjects before treatment, and in 45 major depression patients during treatment with antidepressants. We found that, at baseline, the density (Bmax) of 5-HT2A receptors was significantly higher (by 39%) in depressed patients than in controls. Suicidal patients had significantly higher Bmax values than controls or non-suicidal patients. The rate of serotonin uptake (Vmax), but not the uptake at a single concentration, was significantly higher in depressed patients, particularly in females. There was no significant difference between the Kd or Bmax of [3H]paroxetine binding in control and depressed subjects. Treatment with antidepressant drugs of different pharmacological profile had no significant effect on the density of 5-HT2A receptors, nor did the receptor number predict the response to treatment. The affinity of serotonin uptake site for 5-HT and [3H]paroxetine significantly decreased during treatment with antidepressants, particularly SSRIs. Suppression of 5-HT uptake correlated with decreases in Hamilton depression (HAMD) scores. Our data suggest that the increased density of platelet 5-HT2A receptors may be associated with untreated major depression in antidepressant-free depressed patients, in particular those with suicidal thoughts. The persistence after antidepressant treatment and clinical improvement would suggest that up-regulation of 5-HT2A receptors is a trait rather than state phenomenon. Correlation of 5-HT uptake suppression with decreases in HAMD scores suggests that serotonin uptake inhibition is a relevant factor in antidepressant drug effect and clinical improvement.
Assuntos
Antidepressivos/uso terapêutico , Plaquetas/efeitos dos fármacos , Proteínas de Transporte/efeitos dos fármacos , Transtorno Depressivo/tratamento farmacológico , Receptores de Droga/efeitos dos fármacos , Receptores de Serotonina/efeitos dos fármacos , Serotonina/fisiologia , Adolescente , Adulto , Idoso , Amitriptilina/efeitos adversos , Amitriptilina/uso terapêutico , Antidepressivos/efeitos adversos , Transtorno Depressivo/sangue , Transtorno Depressivo/psicologia , Doxepina/efeitos adversos , Doxepina/uso terapêutico , Feminino , Fluvoxamina/efeitos adversos , Fluvoxamina/uso terapêutico , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Inventário de Personalidade , Receptor 5-HT2A de Serotonina , Trazodona/efeitos adversos , Trazodona/uso terapêutico , Resultado do TratamentoRESUMO
The effects of treatment with serotonin (5-HT) reuptake inhibitors on platelet 5-HT2 receptors, 5-HT reuptake sites an 5-HT uptake were studied in a double-blind trial comparing two selective serotonin reuptake inhibitors (SSRI), paroxetine, and fluoxetine, for the treatment of major depression. Hamilton Depression Rating Scale (HAM-D) scores and platelet 5-HT parameters were determined in 21 depressed patients at baseline, after 4 and 8 weeks of treatment, and were compared to 21 healthy controls. Antidepressant treatment did not significantly alter the density of 5-HT reuptake sites, labelled with [3H]paroxetine, or 5-HT2 receptors, labelled with [3H]LSD. However, a strong correlation was observed between the HAM-D suicidality item and 5-HT2 receptor density at baseline. A marked increase in platelet 5-HT2 receptors at baseline was observed in suicidal depressed patients compared to those with no suicidal ideation and healthy controls. Changes in [3H]paroxetine Bmax and in [3H]5-HT uptake significantly correlated with change in HAM-D score at 4 and 8 weeks respectively. These results confirm previous reports of an association between suicidality and platelet 5-HT2 receptor upregulation. Our data also lends support to the use of platelet 5-HT parameters as indicators of antidepressant efficacy, particularly in suicidal depressed patients.
Assuntos
Plaquetas/efeitos dos fármacos , Transtorno Depressivo/tratamento farmacológico , Fluoxetina/uso terapêutico , Paroxetina/uso terapêutico , Inibidores Seletivos de Recaptação de Serotonina/uso terapêutico , Serotonina/sangue , Adulto , Plaquetas/metabolismo , Transtorno Depressivo/sangue , Método Duplo-Cego , Feminino , Fluoxetina/farmacocinética , Humanos , Dietilamida do Ácido Lisérgico/farmacocinética , Masculino , Pessoa de Meia-Idade , Paroxetina/farmacocinética , Inventário de Personalidade , Receptores de Serotonina/efeitos dos fármacos , Receptores de Serotonina/fisiologia , Inibidores Seletivos de Recaptação de Serotonina/farmacocinéticaRESUMO
Paroxetine is an effective antidepressant drug and potent serotonin (5-HT) uptake inhibitor. It selectively labels 5-HT transporter on platelets and neurons. We report here the synthesis of an aryl-azido derivative of paroxetine, which is a novel photoactive and irreversible ligand for the [3H]paroxetine binding site on the platelet 5-HT transporter. The compound inhibited [3H]paroxetine binding (IC50, 55 nM) and 5-HT uptake (IC50, 12 nM) at equilibrium conditions and inactivated 10-20% of [3H]paroxetine binding sites upon irradiation at 320 nm. SDS-PAGE of platelet protein extract labelled with the radioactive analogue of the synthesized probe revealed the presence of four radioactive bands of which the 71-kDa one was the most prominent.
Assuntos
Marcadores de Afinidade , Proteínas de Transporte/análise , Glicoproteínas de Membrana/análise , Proteínas de Membrana Transportadoras , Proteínas do Tecido Nervoso , Paroxetina/análogos & derivados , Inibidores Seletivos de Recaptação de Serotonina/síntese química , Sítios de Ligação , Plaquetas/metabolismo , Humanos , Cinética , Paroxetina/síntese química , Paroxetina/metabolismo , Fotoquímica , Serotonina/metabolismo , Proteínas da Membrana Plasmática de Transporte de SerotoninaRESUMO
The uptake of [3H]5-HT and the density (Bmax) as well as affinity (Kd) of 5-HT uptake sites labelled with [3H]paroxetine and of 5-HT2 receptors labelled by [3H]LSD were determined in platelets from 25 medication-free patients with major depression and 20 normal controls. The density (Bmax) of 5-HT2 receptors was found to be significantly increased (by 52%) in platelets from depressed patients, particularly females. No changes were found in the affinity (Kd) of 5-HT2 receptors and in 5-HT uptake or [3H]paroxetine binding parameters. Density of 5-HT2 receptors positively correlated with that of [3H]paroxetine sites in control but not in depressed subjects. No correlation was found between the HAMD scores and Bmax of [3H]LSD binding. The results suggest that upregulation of platelet 5-HT2 receptors is a useful biological marker in major depression, particularly in females.
Assuntos
Plaquetas/metabolismo , Transtorno Depressivo/sangue , Receptores de Serotonina/fisiologia , Adolescente , Adulto , Biomarcadores/sangue , Transtorno Depressivo/diagnóstico , Transtorno Depressivo/psicologia , Feminino , Humanos , Dietilamida do Ácido Lisérgico/farmacocinética , Masculino , Pessoa de Meia-Idade , Paroxetina/farmacocinética , Ensaio Radioligante , Receptores de Serotonina/classificação , Fatores Sexuais , Regulação para Cima/fisiologiaRESUMO
Serotonergic function in 22 patients with primary dysthymia and 22 normal volunteers was evaluated by measuring [3H]serotonin uptake and [3H]paroxetine binding in platelets. A significantly lower maximum rate of serotonin uptake was noted in the dysthymic patients than in the normal subjects, indicating a possible serotonergic dysfunction in dysthymia. However, the values for parameters of paroxetine binding were similar in the two groups.
Assuntos
Plaquetas/química , Transtorno Depressivo/sangue , Serotonina/sangue , Adulto , Plaquetas/metabolismo , Transtorno Depressivo/diagnóstico , Transtorno Depressivo/fisiopatologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Modelos Neurológicos , Paroxetina/metabolismo , Receptores de Serotonina/metabolismo , Serotonina/fisiologiaRESUMO
Lectin affinity chromatography was used to demonstrate the glycoprotein nature of the serotonin (5-HT) transporter. The human platelet transporter protein was solubilized with 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS), labelled with [3H]cyanoimipramine and chromatographed on lectin columns. Wheat germ, gorse seed and lentil lectin-agarose columns specifically retained 5-HT transporter. Peanut and horse gram lectins were ineffective. Concanavalin A showed high non-specific adsorption. Binding of [3H]imipramine to platelet membranes or solubilized 5-HT transporter was not affected by lectins. These data suggest that the lectin interaction with 5-HT transporter is extrinsic to the antidepressant binding site.
Assuntos
Plaquetas/metabolismo , Proteínas de Transporte/metabolismo , Lectinas/farmacologia , Glicoproteínas de Membrana/metabolismo , Proteínas de Membrana Transportadoras , Proteínas do Tecido Nervoso , Serotonina/metabolismo , Sítios de Ligação , Proteínas de Transporte/química , Ácidos Cólicos , Cromatografia de Afinidade , Detergentes , Humanos , Imipramina/metabolismo , Lectinas/metabolismo , Glicoproteínas de Membrana/química , Proteínas da Membrana Plasmática de Transporte de SerotoninaRESUMO
Experimental evidence suggests the occurrence of endogenous antidepressant-like compounds in the brain, blood plasma, and urine. After extensive purification by calmodulin-sepharose affinity chromatography, and further purification of the urine-derived sample by exclusion chromatography, at least three distinctive fractions have been identified. These fractions effectively inhibited serotonin uptake, imipramine, and/or paroxetine binding, and they apparently contained some compounds that were recognized by the anti-imipramine or anti-paroxetine antibodies. Their identification may have significant implications for studies of affective illness.
Assuntos
Fatores Biológicos/metabolismo , Inibidores Seletivos de Recaptação de Serotonina , Serotonina/metabolismo , Adulto , Anticorpos/análise , Fatores Biológicos/análise , Encéfalo/metabolismo , Calmodulina/metabolismo , Cromatografia em Agarose , Cromatografia em Gel , Transtorno Depressivo/tratamento farmacológico , Transtorno Depressivo/imunologia , Transtorno Depressivo/metabolismo , Feminino , Humanos , Imipramina/imunologia , Imipramina/metabolismo , Imipramina/uso terapêutico , Masculino , Paroxetina/imunologia , Paroxetina/metabolismo , Paroxetina/uso terapêutico , Radioimunoensaio , Serotonina/imunologia , Serotonina/uso terapêutico , Relação Estrutura-AtividadeRESUMO
Calf brain and human platelet extracts purified by Bio-Gel P2 column chromatography contained substances that inhibited serotonin uptake and 3H-imipramine binding. Some of these endogenous substances were also recognized by rabbit antibodies produced against imipramine. The data suggest the possible existence of endogenous serotonin uptake modulators, which may possess a partial molecular structure similar to that identified by the antibodies.
Assuntos
Anticorpos , Plaquetas/metabolismo , Proteínas de Transporte , Lobo Frontal/metabolismo , Imipramina/imunologia , Receptores de Droga , Receptores de Neurotransmissores/antagonistas & inibidores , Antagonistas da Serotonina , Serotonina/sangue , Animais , Bovinos , Humanos , Imipramina/farmacocinética , Ensaio Radioligante , Receptores de Neurotransmissores/metabolismo , Receptores de Serotonina/metabolismoRESUMO
Inhibition of high-affinity [3H]-imipramine binding to platelet membranes by human plasma fractions and isolated plasma proteins was investigated. Several plasma proteins were found to contribute to the observed apparent inhibition and this contribution was assessed in terms of inhibitor units. Alpha 1 acid glycoprotein, high density and low density lipoprotein, IgG and alpha 1-antitrypsin were identified as effective non-specific inhibitors. Alpha-1-acid glycoprotein was confirmed to be the most potent plasma protein inhibitor. Cohn fractions were evaluated for the presence of the postulated endocoid of [3H]-imipramine binding site.
Assuntos
Plaquetas/metabolismo , Proteínas Sanguíneas/farmacologia , Imipramina/sangue , Sítios de Ligação , Membrana Celular/metabolismo , Eletroforese em Gel de Poliacrilamida , Humanos , Serotonina/metabolismo , TrítioRESUMO
The effect of chlorpromazine on subcellular biosynthesis, hydrolysis, and transfer of lipids and liponucleotides participating in the biosynthesis of polyglycerophosphatides in guinea pig liver was studied. Chlorpromazine showed an apparent stimulation of accumulation of phosphatidic acid and CDP-diglycerides in microsomal membranes and phosphatidylglycerolphosphate in mitochondrial membranes in a concentration-dependent manner that was influenced by incubation time and the nature of fatty acids in CDP-diglycerides. Transfer of membrane-bound CDP-diglycerides from microsomal to mitochondrial membranes was established by the CDP-diglyceride-dependent biosynthesis of phosphatidylglycerolphosphate and phosphatidylglycerol and appeared to be inhibited by the addition of chlorpromazine by about 20%. Evidence was obtained for the formation of a molecular complex between phosphatidic acid and chlorpromazine; this was thought to be responsible for the protection from phosphatidate phosphohydrolase at the concentrations of chlorpromazine and Mg2+ examined.
Assuntos
Clorpromazina/farmacologia , Nucleotídeos de Citosina/metabolismo , Metabolismo dos Lipídeos , Microssomos Hepáticos/metabolismo , Mitocôndrias Hepáticas/metabolismo , Fosfatidilgliceróis/biossíntese , Animais , Cardiolipinas/biossíntese , Clorpromazina/metabolismo , Diglicerídeos de Citidina Difosfato/biossíntese , Cobaias , Hidrólise , Lipossomos/metabolismo , Microssomos Hepáticos/efeitos dos fármacos , Mitocôndrias Hepáticas/efeitos dos fármacos , Ácidos Fosfatídicos/biossíntese , Ácidos Fosfatídicos/metabolismoRESUMO
Evidence for an association of both hydrophilic insulin and hydrophobic cyclosporin with liposomes prepared from egg yolk lecithin, cholesterol, and stearylamine (7:2:2.25 molar ratio) was obtained by Sepharose-4B gel filtration. The method used to prepare unilamellar liposomes containing 29.7 nmol cyclosporin and 2.3 nmol insulin per mu mol of liposomal lecithin is described.
Assuntos
Ciclosporinas , Insulina , Lipossomos , Fosfatidilcolinas , 1,2-Dipalmitoilfosfatidilcolina , Aminas , Radioisótopos de Carbono , Colesterol , MétodosRESUMO
Small unilamellar [14C]lecithin liposomes prepared in the presence of cyclosporin sedimented at 12,000 g. Sepharose 4B gel filtration of the resuspended pellet and supernatant yielded identical peaks consisting of small unilamellar liposomes containing cyclosporin. The column retained 40-50 per cent of 14C-labelled liposomes prepared in the presence of cyclosporin as liposome aggregates.
Assuntos
Ciclosporinas/administração & dosagem , Ciclosporinas/metabolismo , Portadores de Fármacos , LipossomosAssuntos
Ciclosporinas/administração & dosagem , Lipossomos , Colesterol , Fosfatidilinositóis , ÁguaRESUMO
Participation of microsomal CDP-diglycerides in mitochondrial biosynthesis of phosphatidylglycerol was studied by [3H]palmitoyl, [14C]linoleoyl, and [14C]arachidonoyl CDP-diglycerides and [3H]CDP-diglycerides which were bound to microsomal membranes, incubated with unlabelled mitochondrial membranes, and further incubated in the presence of radioactive sn-glycero-3-phosphate under conditions required for mitochondrial phosphatidylglycerol biosynthesis. Ten to 15% of microsomal radioactive CDP-diglycerides was transferred to mitochondrial membranes and incorporated into mitochondrial radioactive lipids identified as phosphatidylglycerol, phosphatidylglycerophosphate, and, when [14C]linoleoyl CDP-diglycerides were used, diphosphatidylglycerol (cardiolipin).
Assuntos
Diglicerídeos de Citidina Difosfato/metabolismo , Microssomos Hepáticos/metabolismo , Mitocôndrias Hepáticas/metabolismo , Açúcares de Nucleosídeo Difosfato/metabolismo , Fosfatidilgliceróis/biossíntese , Animais , Radioisótopos de Carbono , Cobaias , Membranas Intracelulares/metabolismo , Cinética , TrítioRESUMO
Basic substances and acidic ionophores that increase the lysosomal pH in cultured macrophages (Ohkuma, S., and B. Poole, 1978, Proc. Natl. Acad. Sci. USA., 75:3327-3331; Poole, B., and S. Ohkuma, 1981, J. Cell Biol., 90:665-669) inhibited the digestion of heat-denatured acetylated bovine serum albumin (BSA) taken up by the cells. For several substances, the shift in pH sufficed to explain the inhibition of proteolysis. Additional effects, presumably on enzyme activities, have to be postulated for tributylamine, amantadine, and chloroquine. Sodium fluoride (10 mM) had no significant effect on the breakdown of BSA by macrophages. The breakdown of endogenous macrophage proteins, whether short lived or long lived, was inhibited approximately 40% by 10 mM NaF and 30%, or sometimes less in the case of long-lived proteins, by 100 microM chloroquine. When the cells were supplied with BSA, a mixture of cell proteins, or even inert endocytosible materials, the breakdown of endogenous long-lived proteins and the inhibitory effect of chloroquine on this process were selectively reduced. Inhibition of endocytosis by cytochalasins B or D did not affect the chloroquine-sensitive breakdown of endogenous proteins, indicating that the proteins degraded by this process were truly endogenous and not taken in from the outside by cellular cannibalism. On the other hand, when macrophage proteins were supplied extracellularly, their breakdown occurred at the same rate for short-lived and long-lived proteins, and it was strongly inhibited by chloroquine and not by NaF. It is concluded from these results that the breakdown of endogenous proteins, both short-lived and long-lived, probably takes place partly (approximately 30%) in lysosomes and partly through one or more nonlysosomal mechanism(s) unaffected by chloroquine and presumably susceptible to inhibition by fluoride. A difference must exist between short-lived and long-lived proteins in the manner in which they reach lysosomes or are handled by these organelles; this difference would account for the selective effect of the supply of endocytosible materials on the lysosomal processing of long-lived proteins.
Assuntos
Cloroquina/farmacologia , Macrófagos/metabolismo , Proteínas/metabolismo , Fluoreto de Sódio/farmacologia , Animais , Bovinos , Células Cultivadas , Citocalasinas/farmacologia , Depressão Química , Endocitose/efeitos dos fármacos , Concentração de Íons de Hidrogênio , Lisossomos/metabolismo , Macrófagos/efeitos dos fármacos , Camundongos , Cavidade Peritoneal/citologia , Soroalbumina Bovina/metabolismoRESUMO
The study examined the effect of an association of chlorpromazine, phosphatidic acid and cytidine-diphosphoryl-1,2-diglycerides (CDP-diglycerides) with small unilamellar lecithin liposomes on the formation, hydrolysis and transfer of lipids and cytidine liponucleotides in microsomal and mitochondrial membranes isolated from guinea-pig liver. Association with liposomes undermined the effect of chlorpromazine on these processes, but the type of effect, i.e. inhibitory or stimulatory, was retained. Association of CDP-diglycerides with small unilamellar lecithin liposomes tended to protect this substrate from subcellular uptake, thereby inhibiting phosphatidylinositol and polyglycerophosphatide formation. Phosphatidic acid in the form of liposomes stimulated CDP-diglyceride formation. The nature of fatty acids influenced the magnitude of these effects in polyglycerophosphatide biosynthesis. Transfer of CDP-diglycerides from microsomal to mitochondrial membranes was inhibited by both chlorpromazine associated with liposomes and liposomes alone.
Assuntos
Clorpromazina/administração & dosagem , Fígado/metabolismo , Lipídeos de Membrana/biossíntese , Animais , Clorpromazina/farmacologia , Portadores de Fármacos , Cobaias , Lipossomos , Fígado/efeitos dos fármacosRESUMO
The mechanism of the previously reported cytotoxicity of liposomes containing plant phosphatidylinositol (PI) against numerous tumor cell lines was examined in detail by using liposomes containing synthetic PI specifically labeled either with radioactive myo-inositol, or in the sn-2 position with radioactive linoleic acid, oleic acid, or arachidonic acid. The uptake of liposomal PI by N4TG1 neuroblastoma cells increased with time and was dependent on the nature of the fatty acids. Uptake was highest with liposomal PI containing linoleic acid followed by arachidonic acid and then by oleic acid. The cellular fate of liposomal PI was determined by analysis of radioactive metabolites present in extracts of tumor cell lipids. Appearance of liposomal PI metabolic products in the tumor cells was correlated with thymidine uptake as a measure of viability. After 3 h incubation of cells with PI liposomes it was found that the release of both radioactive liposomal fatty acids (and probably also lyso-Pl) and radioactive diglycerides was correlated inversely with the cellular uptake of [methyl-3H]thymidine and uptake of [3H]myoinositol. An experiment in which liposomes were prepared both from animal Pl which contained predominantly saturated fatty acids in the sn-2 position and an increasing mole fraction of a synthetic Pl containing radioactive linoleic acid in the sn-2 position established that the amount of Pl containing linoleic acid in the sn-2 position could be correlated with a decrease in the amount of thymidine uptake by tumor cells. The above results clearly established that phospholipases A2 and C in the tumor cells were responsible for the formation of metabolites of liposomal Pl, and these metabolic products might have been responsible for cytotoxicity and cell death.
Assuntos
Lipossomos/metabolismo , Lisofosfolipídeos , Neoplasias Experimentais/metabolismo , Fosfatidilinositóis/metabolismo , Animais , Linhagem Celular , Ácido Linoleico , Ácidos Linoleicos/metabolismo , Camundongos , Neoplasias Experimentais/patologia , Fosfatidilinositóis/farmacologiaRESUMO
The effect of adding small unilamellar lecithin liposomes, prepared in the presence of cytidine-diphosphoryl-1,2-diglycerides (CDP-diglycerides) or cytochrome c, on microsomal biosynthesis of phosphatidylinositol and NADPH-cytochrome c reduction and on mitochondrial biosynthesis of polyglycerophosphatides and succinate-cytochrome c reduction was studied in isolated guinea-pig liver subcellular membranes. Both microsomal biosynthesis of phosphatidylinositol and mitochondrial biosynthesis of phosphatidylglycerol were significantly reduced when CDP-diglycerides associated with liposomes were used, suggesting that some CDP-diglycerides were entrapped by liposomal membranes and were not available to subcellular membranes as substrates. The degree of decrease in phospholipid biosynthesis depended on the membrane and the nature of fatty acids in CDP-diglycerides. The composition of mitochondrial polyglycerophosphatides synthesized in the presence of CDP-diglycerides-liposomes was also affected in respect to the amount of phosphatidylglycerol formed. The reduction of cytochrome c in both microsomal and mitochondrial membranes was also decreased when liposomes were present in the assay system, but to a lesser degree than the phospholipid biosynthesis. These results indicate that the cytochrome c liposome association did not provide efficient protection of this substrate from the subcellular reduction. When chlorpromazine was also present with liposomes in the assay system, the NADPH-cytochrome c reduction in microsomes was scarcely affected, while the succinate-cytochrome c reduction in mitochondria was dependent on the concentration of chlorpromazine and could be completely abolished. These results were interpreted in terms of liposomal interaction with substrates in competition with subcellular membranes for the same substrates.
Assuntos
Lipossomos/metabolismo , Microssomos Hepáticos/metabolismo , Mitocôndrias Hepáticas/metabolismo , Animais , Grupo dos Citocromos c/metabolismo , Cobaias , Técnicas In Vitro , Oxirredução , Fosfatidilgliceróis/biossíntese , Fosfatidilinositóis/biossínteseRESUMO
Transfer of membrane-bound phosphatidyl-[2'-3H]inositol from microsomal to unlabelled mitochondrial and from mitochondrial to unlabelled microsomal membranes was studied using partially purified cytosol proteins isolated from guinea pig liver cytosol. In the absence and presence of these proteins the amounts of phosphatidylinositol transfer from microsomal to mitochondrial membranes were approximately 21 and 33%, respectively, and the amounts from mitochondrial to microsomal membranes were approximately 31 and 39%, respectively. The release of phosphatidyl-[2'-3H]inositol from microsomal membranes in the absence of mitochondria was dependent on concentration of cytosol proteins. Two mechanisms for movement between membranes are proposed. In cytosol-protein-independent movement of phosphatidyl-[2'-3H]inositol from microsomal to mitochondrial membranes, a direct contact between membranes is required, since phosphatidyl-[2'-3H]inositol was not detected in the reaction medium. In the cytosol-protein-catalyzed transfer, formation of phosphatidyl-[2'-3H]inositol - cytosol protein complex is postulated, since phosphatidyl-[2'-3H]inositol was released into the reaction medium and its movement proceeded from mitochondrial to microsomal membranes in the presence of partially purified cytosol proteins. Thus, contact between the two membranes is probably not necessary for this transfer. Implications for the movement of phospholipids between biological membranes are discussed.