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1.
Kinetic analysis of 3H-serotonin accumulation by crude synaptosomal suspensions of neocortex, hippocampus and caudate or by whole homogenates of cerebellum revealed the presence of a high affinity uptake component having an apparent Km for serotonin which ranged from 2.8 to 6.0 × 10?8 M. A second, low affinity, uptake component with an apparent Km of 7 × 10?6 M was present in caudate. A comparable low affinity uptake component for serotonin was not observed in neocortex, hippocampus or cerebellum. Lesions in the medial forebrain bundle produced significant decreases in serotonin comtent of neocortes, hippocampus and caudate (66 to 75%) and a significant increase in serotonin content of cerebellum (25%). The lesions did not affect the apparent Km of the high affinity uptake system but did produce change in Vmax which paralleled the changes in content of serotonin. The lesions also produced decreases in dopamine and norepinephrine content of caudate and a comparable decrease in the Vmax of the low affinity uptake system with no change in the apparent Km. There was a correlation of 0.97 between the endogenous content of serotonin and the Vmax of the high affinity uptake system. These results support the view that the high and low affinity components of serotonin uptake represent accumulation into serotonergic and catecholaminergic neurons, respectively. 相似文献
2.
《Nucleosides, nucleotides & nucleic acids》2013,32(4-7):343-360
Compound 20 (CVT-3146 - a 2-[(N-1-(4-N-methylcarboxamidopyrazolyl)] adenosine derivative) and compound 31 (CVT-3033 - a 2-[(4-(1-N-pentyl-pyrazolyl)] adenosine derivative), were found to be short acting functionally selective coronary vasodilators (CV t0.5 = 5.2 ± 0.2 and 3.4 ± 0.5 min, respectively - rat isolated heart 50% reversal time) with good potency (EC50s = 6.4 ± 1.2 nM and 67.9 ± 16.7 nM, respectively), but they possess low affinity for the ADO A2A receptor (Ki = 1122 ± 323 nM and 2138 ± 952 nM, respectively; pig striatum). 相似文献
3.
Georg Sager Natalia Smaglyukova Ole‐Martin Fuskevaag 《Journal of cellular physiology》2018,233(8):5972-5980
The present study was conducted to characterise the transporter(s) responsible for the uptake of cyclic nucleotides to human erythrocytes. Western blotting showed that hRBC expressed OAT2 (SLC22A7), but detection of OAT1 (SLC22A6), or OAT3 (SLC22A8) was not possible. Intact hRBC were employed to clarify the simultaneous cyclic nucleotide egression and uptake. Both these opposing processes were studied. The Km‐values for high affinity efflux was 3.5 ± 0.1 and 39.4 ± 5.7 μM for cGMP and cAMP, respectively. The respective values for low affinity efflux were 212 ± 11 and 339 ± 42 μM. The uptake was characterised with apparently low affinity and similar Km‐values for cGMP (2.2 mM) and cAMP (0.89 mM). Using an iterative approach in order to balance uptake with efflux, the predicted real Km‐values for uptake were 100–200 μM for cGMP and 50–150 μM for cAMP. The established OAT2‐substrate indomethacin showed a competitive interaction with cyclic nucleotide uptake. Creatinine, also an OAT2 substrate, showed saturable uptake with a Km of 854 ± 98 μM. Unexpectedly, co‐incubation with cyclic nucleotides showed an uncompetitive inhibition. The observed Km‐values were 399 ± 44 and 259 ± 30 μM for creatinine, in the presence of cGMP and cAMP, respectively. Finally, the OAT1‐substrate para‐aminohippurate (PAH) showed some uptake (Km‐value of 2.0 ± 0.4 mM) but did not interact with cyclic nucleotide or indomethacin transport. 相似文献
4.
Saturable, Energy-Dependent Uptake of Phenanthrene in Aqueous Phase by Mycobacterium sp. Strain RJGII-135
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Naoyuki Miyata Keisuke Iwahori Julia M. Foght Murray R. Gray 《Applied microbiology》2004,70(1):363-369
The mechanism of uptake of phenanthrene by Mycobacterium sp. strain RJGII-135, a polycyclic hydrocarbon-degrading bacterium, was examined with cultures grown on phenanthrene (induced for phenanthrene metabolism) and acetate (uninduced). Washed cells were suspended in aqueous solutions of [9-14C]phenanthrene, and then the cells were collected by filtration. Low-level steady-state 14C concentrations in uninduced cells were achieved within the first 15 s of incubation. This immediate uptake did not show saturation kinetics and was not susceptible to inhibitors of active transport, cyanide and carbonyl cyanide m-chlorophenylhydrazone. These results indicated that phenanthrene enters rapidly into the cells by passive diffusion. However, induced cells showed cumulative uptake over several minutes. The initial uptake rates followed saturation kinetics, with an apparent affinity constant (Kt) of 26 ± 3 nM (mean ± standard deviation). Uptake of phenanthrene by induced cells was strongly inhibited by the inhibitors. Analysis of cell-associated 14C-labeled compounds revealed that the concurrent metabolism during uptake was rapid and was not saturated at the substrate concentrations tested, suggesting that the saturable uptake observed reflects membrane transport rather than intracellular metabolism. These results were consistent with the presence of a saturable, energy-dependent mechanism for transport of phenanthrene in induced cells. Moreover, the kinetic data for the cumulative uptake suggested that phenanthrene is specifically bound by induced cells, based on its saturation with an apparent dissociation constant (Kd) of 41 ± 21 nM (mean ± standard deviation). Given the low values of Kt and Kd, Mycobacterium sp. strain RJGII-135 may use a high-affinity transport system(s) to take up phenanthrene from the aqueous phase. 相似文献
5.
We have investigated alterations in beta-adrenergic receptors in rat myocardial membranes derived from hypothyroid and hyperthyroid animals. (-)Isoproterenol competition curves with (-)[3H]dihydroalprenolol revealed that isoproterenol binds to the beta-adrenergic receptor with two distinct affinity states having high (KH) and low (KL) dissociation constants. In the presence of guanine nucleotides the isoproterenol competition curve steepened and had a higher EC50 (50% displacement). This was due to a transition of the high affinity state to a uniformly low affinity state. Using computer modeling of these competition curves, we have demonstrated that in hyperthyroidism, the isoproterenol curve in the absence of guanine nucleotides is shifted to the left with the EC50 changing from 180 ± 40 to 80 ± 20 nM (p < .02). The fold shift (4 fold) in KH (nM) 30 ± 9 to 7 ± 2 (p < .001) is greater than that (1.6 fold) in KL (nM) 595 ± 56 to 376 ± 34 (p < .001) such that the KL/KH ratio shifted from 20 ± 3 to 54 ± 9 (p < .001). The ratio, KL/KH, for a particular agonist appears to be related to its efficacy in activating adenylate cyclase.There was no significant alteration in any of these parameters in hypothyroid animals. Receptor number was decreased in hypothyroidism, 16 ± 3 fmol/mg protein (p < .03) and increased in hyperthyroidism 44 ± 4 (p < .03) compared to control 26 ± 2.In the rat heart agonist affinity and receptor number are modulated in hyperthyroidism, but only receptor number in hypothryoidism. Thus thyroid hormone can modify not only receptor number but agonist affinity as well. 相似文献
6.
The characteristics of malate transport into aerobically grown cells of the purple photosynthetic bacterium Rhodobacter capsulatus were determined. A single transport system was distinguished kinetically which displayed a Kt value of 2.9 ± 1.2 μM and Vmax of 43 ± 6 nmol · min-1 · mg-1 protein. Competition experiments indicated that the metabolically related C4-dicarboxylates succinate and fumarate are also
transported by this system. Malate uptake was sensitive to osmotic shock and evidence from the binding of radiolabelled malate
and succinate to periplasmic protein fractions indicated that transport is mediated by a dicarboxylate binding protein. The
activity of the transport system was studied as a function of external and internal pH and it was found that a marked activation
of uptake occurred at intracellular pH values greater than 7. The use of a high affinity binding protein dependent system
to transport a major carbon and energy source suggests that Rhodobacter capsulatus would be capable of obtaining growth sustaining quantities of C4-dicarboxylates even if these were present at very low concentrations
in the environment. 相似文献
7.
Kannan Subramanian Conan J. Fee Rayleen Fredericks Richard S. Stubbs Mark T. Hayes 《Journal of molecular recognition : JMR》2013,26(12):643-652
Type 2 diabetes affects millions of people worldwide, and measuring the kinetics of insulin receptor‐insulin interactions is critical to improving our understanding of this disease. In this paper, we describe, for the first time, a rapid, real‐time, multiplex surface plasmon resonance (SPR) assay for studying the interaction between insulin and the insulin receptor ectodomain, isoform A (eIR‐A). We used a scaffold approach in which anti‐insulin receptor monoclonal antibody 83–7 (Abcam, Cambridge, UK) was first immobilized on the SPR sensorchip by amine coupling, followed by eIR‐A capture. The multiplex SPR system (ProteOn XPR36?, Bio‐Rad Laboratories, Hercules, CA) enabled measurement of replicate interactions with a single, parallel set of analyte injections, whereas repeated regeneration of the scaffold between measurements caused variable loss of antibody activity. Interactions between recombinant human insulin followed a two‐site binding pattern, consistent with the literature, with a high‐affinity site (dissociation constant KD1 = 38.1 ± 0.9 nM) and a low‐affinity site (KD2 = 166.3 ± 7.3 nM). The predominantly monomeric insulin analogue Lispro had corresponding dissociation constants KD1 = 73.2 ± 1.8 nM and KD2 = 148.9 ± 6.1 nM, but the fit to kinetic data was improved when we included a conformational change factor in which the high‐affinity site was converted to the low‐affinity site. The new SPR assay enables insulin‐eIR‐A interactions to be followed in real time and could potentially be extended to study the effects of humoral factors on the interaction, without the need for insulin labeling. Copyright © 2013 John Wiley & Sons, Ltd. 相似文献
8.
Jian Guo Amber L. Annett Rebecca L. Taylor Suzanne Lapi Thomas J. Ruth Maria T. Maldonado 《Journal of phycology》2010,46(6):1218-1228
We investigated copper (Cu) acquisition mechanisms and uptake kinetics of the marine diatoms Thalassiosira oceanica Hasle, an oceanic strain, and Thalassiosira pseudonana Hasle et Heimdal, a coastal strain, grown under replete and limiting iron (Fe) and Cu availabilities. The Cu‐uptake kinetics of these two diatoms followed classical Michaelis–Menten kinetics. Biphasic uptake kinetics as a function of Cu concentration were observed, suggesting the presence of both high‐ and low‐affinity Cu‐transport systems. The half‐saturation constants (Km) and the maximum Cu‐uptake rates (Vmax) of the high‐affinity Cu‐transport systems (~7–350 nM and 1.5–17 zmol · μm?2 · h?1, respectively) were significantly lower than those of the low‐affinity systems (>800 nM and 30–250 zmol · μm?2 · h?1, respectively). The two Cu‐transport systems were controlled differently by low Fe and/or Cu. The high‐affinity Cu‐transport system of both diatoms was down‐regulated under Fe limitation. Under optimal‐Fe and low‐Cu growth conditions, the Km of the high‐affinity transport system of T. oceanica was lower (7.3 nM) than that of T. pseudonana (373 nM), indicating that T. oceanica had a better ability to acquire Cu at subsaturating concentrations. When Fe was sufficient, the low‐affinity Cu‐transport system of T. oceanica saturated at 2,000 nM Cu, while that of T. pseudonana did not saturate, indicating different Cu‐transport regulation by these two diatoms. Using CuEDTA as a model organic complex, our results also suggest that diatoms might be able to access Cu bound within organic Cu complexes. 相似文献
9.
《Neurochemistry international》1987,11(1):89-93
The uptake and accumulation of N-methyl-4-phenylpyridinium ion (MPP+), a neurotoxin produced by oxidation of N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), into PC12h pheochromocytoma cells were examined. Concentration gradients of MPP+ were established at its low concentrations of 10 to 100 nM. Uptake of MPP+ into PC12h cells was mediated by saturable, carrier mediated transport systems with two different kinetic properties; a high-affinity and low-capacity system and a low-affinity and high-capacity system. The apparent Km values of these two systems were obtained to be 254.4 ± 96.5 nM and 23.1 ± 6.9 μM, respectively, and the maximal uptake velocity was obtained to be 8.47 ± 1.72 and 28.6 ± 5.2 pmol/min/mg protein, respectively. The uptake by a high-affinity system was mediated by a carrier system common to dopamine and noradrenalin and MPTP itself proved to be taken up by this system, which was further confirmed by the inhibition of the MPP+ uptake by nomifensine and mazindol. The uptake was inhibited by metabolic inhibitors, such as carbonyl cyanide m-chlorophenyl hydrazone, sodium cyanide and 2,4-dinitrophenol, and the uptake was inhibited by ouabain and nigercin. By subcellular fractionation, MPP+ taken up was found to be localized mainly in cytosol fraction, but a definite amount of MPP+ was found also in mitochondrial fraction. 相似文献
10.
Two sets of high-affinity thyroxine binding sites (KD 0.39 ± 0.06 nM and 23 ± 5 nM) were detected on purified rat liver plasma membranes. Thyroxine is bound with high stereospecificity regarding iodine substituents and alanine side chain modifications of the molecule. Thyroxine binding is inhibited by -SH blocking agents and proteases. The highest affinity thyroxine binding site is also affected by phospholipase A and is distinct from triiodothyronine binding sites present in the membrane preparations; arguments are given for its plasmalemma origin. 相似文献
11.
The oligopeptide chemotactic factor receptor on human polymorphonuclear leukocyte membranes exists in two affinity states 总被引:14,自引:0,他引:14
C Koo R J Lefkowitz R Snyderman 《Biochemical and biophysical research communications》1982,106(2):442-449
The binding of the chemoattractant N-formyl-methionylleucyl-[3H]phenylalanine to intact polymorphonuclear leukocytes and membrane preparations was analyzed by computer methods. Whole viable cells bind the chemoattractant with a single dissociation constant (KD) of 22.3 ± 2.4 nM and contain an average of 55,000 receptors percell. In contrast, the binding data using membrane preparations were consistent with the presence of two classes of binding sites with average KDs of 0.53 ± 0.01 nM and 24.4 ± 1.2 nM. The high affinity receptors accounted for ca. 25% of the binding sites. Increasing the receptor occupancy did not affect the rate of dissociation of the ligand-receptor complex thus negative cooperativity is not a likely explanation for the complex binding isotherms. On the other hand, the dissociation kinetics did agree with the two affinity receptor model. 相似文献
12.
《Journal of receptor and signal transduction research》2013,33(5):653-666
AbstractSpecific binding sites for somatostatin have been detected in cytosolic fraction of bovine cystic duct mucosa. At 37°C, the interaction of 125I-Tyr11-somatostatin with cytosolic fraction was rapid, reversible, specific and saturable. At equilibrium, the binding of tracer was competitively inhibited by native peptide in the 1 nM to 2 µ M range of concentrations. Scatchard analysis of binding data suggested the presence of two distinct classes of somatostatin binding sites: a class with a high affinity (Kd = 7.8 ± 0.3 nM) and a low capacity (1.3 ± 0.3 pmol somatostatin/mg protein) and a class with a low affinity (Kd = 129.1 ± 2.0 nM) and a high capacity (43.5 ± 6.7 pmol somatostatin/mg protein). The binding sites were shown to be highly specific for somatostatin since neuropeptides present in cystic duct such as Leu-enkephalin, neurotensin, substance P and vasoactive intestinal peptide did practically not show competition. These findings suggest that somatostatin could contribute to the regulation of the functions of the cystic duct mucosa in physiological and pathological conditions. 相似文献
13.
A selective agonist of non-opioid β-endorphin receptor synthetic peptide octarphin (TPLVTLFK, specific activity 28 Ci/mmol)
was prepared. The [3H]octarphin binding to rat myocardium membranes before and after experimental myocardial infarction (EMI) was studied. It
was found that [3H]octarphin with high affinity and specificity binds to non-opioid β-endorphin receptor of rat myocardium membranes before
EMI: K
d1 value of the [3H]octarphin specific binding to membranes was 1.8 ± 0.2 nM. In 3 h after EMI a sharp lowering in affinity of the binding is
observed (K
d2 = 13.3 ± 0.4 nM), and in 48 h its almost complete restoration (K
d4 = 2.2 ± 0.3 nM). The results indicate participation of non-opioid β-endorphin receptor in the regulation of myocardial activity. 相似文献
14.
Five catechol estrogens and two 2-methoxyestrogens were compared for their relative affinity of binding to hypothalamic, pituitary and uterine cytosol estrogen receptors; and for the kinetics of the catechols' methylation by hepatic catechol-O-methyltransferase. All of the catechol estrogens tested have similar Km 's for O-methylation (9–14 μM). Estrogen receptor affinities, however, differ widely. In hypothalamus, for example, where estradiol-17β has a Kd of 0.039 ± 0.008 nanomolar, 4-hydroxyestradiol also binds tightly (0.12 ± 0.02 nM), 2-hydroxyestradiol and 4-hydroxyestrone with intermediate affinity (0.26 ± 0.06 and 0.28 ± 0.07 nM, respectively), and 2-hydroxyestrone and 2-hydroxyestriol much less well (1.68 ± 0.79 and 1.27 ± 0.26 nM, respectively). The binding of the 2-methoxyestrogens is extremely weak. These receptor affinities roughly parallel the potencies of these compounds in altering gonadotropin secretion. 相似文献
15.
Parham Taslimi Sabiya Osmanova İlhami Gulçin Sabira Sardarova Vagif Farzaliyev Afsun Sujayev Ruya Kaya Fatma Koc Sukru Beydemir Saleh H. Alwasel Omer Irfan Kufrevioglu 《Journal of biochemical and molecular toxicology》2017,31(9)
Compounds containing nitrogen and sulfur atoms can be widely used in various fields, including industry, medicine, biotechnology, and chemical technology. Among them, amides of acids and heterocyclic compounds have an important place. These amides and thiazolidine‐4‐ones showed good inhibitory action against butyrylcholinesterase (BChE), acetylcholinesterase (AChE), and human carbonic anhydrase isoforms. AChE exists at high concentrations in the brain and red blood cells. BChE is an important enzyme that is plentiful in the liver, and it is released into the blood in a soluble form. They were demonstrated to have effective inhibition profiles with Ki values of 23.76–102.75 nM against hCA I, 58.92–136.64 nM against hCA II, 1.40–12.86 nM against AChE, and 9.82–52.77 nM against BChE. On the other hand, acetazolamide showed Ki value of 482.63 ± 56.20 nM against hCA I, and 1019.60 ± 163.70 nM against hCA II. Additionally, Tacrine inhibited AChE and BChE, showing Ki values of 397.03 ± 31.66 and 210.21 ± 15.98 nM, respectively. 相似文献
16.
《Journal of receptor and signal transduction research》2013,33(2-3):121-131
AbstractThe fluorescent ß-adrenergic receptor probe alprenolol-NBD was found to exhibit a high affinity (Kd 3.2 nM) and a low capacity (10 fmol/mg protein) for the ß2-adrenergic receptor on living Chang liver cells but also a high affinity (Kd 320 nM) for non-ß-adrenergic receptor binding sites with a very high capacity (28,000 fmol/mg protein). Calculations are presented which make clear that less than 3% of the binding of alprenolol-NBD during visualization experiments is ß-adrenergic receptor related. Furthermore, it is shown that besides the downregulation of ß-adrenergic receptors during incubation with isoproterenol, the high-affinity non-ß-receptor binding sites are also deminishing during incubation with isoproterenol. Based on our findings it is concluded that the results of Henis et al. (1) who claimed the visualization of the ß-adrenergic receptor population on Chang liver cells by alprenolol-NBD must be interpreted as an almost completely non-specific fluorescence. 相似文献
17.
《Journal of receptor and signal transduction research》2013,33(1):95-106
AbstractMesenteric artery smooth muscle cells were grown in culture media containing high, normal, or low concentrations of potassium to study the effects on angiotensin II (Ang II) receptor regulation. Cell growth was similar among cells grown in the different culture media. Cells grown in high potassium media (K=5.8 mEq/L) had an equilibrium dissociation constant, Kd, of 1.59 ± 0.2 nM, whereas those grown in normal potassium media (K=4.1 mEq/L) had a Kd of 1.79 ± 0.2 nM and those grown in a low potassium media (K=2.9 mEq/L) had a Kd of 1.19 ± 0.12 nM (not significantly different, NS). Binding capacity of smooth muscle cells grown in high potassium media was 81 ± 16.7 fmol/mg prot, 95.1 ± 12.4 fmol/mg prot in those grown in normal potassium media and those grown in low potassium media 86.4 ± 24.1 fmol/mg prot (NS). Binding of radiolabelled Ang II was reduced by approximately 70% in cells exposed to unlabelled Ang II for 30 or 60 minutes. However, this effect of exposure to Ang II to reduce subsequent binding of Ang II was identical in cells grown in high and low potassium medium. Therefore, we were unable to identify a direct effect of low potassium to induce changes in Ang II receptor binding affinity or binding capacity. Previously observed changes in these Ang II binding parameters in potassium-depleted rats was probably a consequence of other factors which were simultaneously altered by potassium deficiency. 相似文献
18.
Lorne J. Brandes Gary M. Queen Frank S. LaBella 《Journal of cellular biochemistry》1998,69(3):233-243
Histamine and polyamines have been implicated in the mediation of cell proliferation. Our previous work linked the growth-modulatory effects of histamine with its binding to intracellular sites in microsomes and nuclei of various tissues. In this study, we identify cytochrome P450 enzymes as a major component of microsomal intracellular sites in hepatocytes and demonstrate that polyamines compete with high affinity for histamine binding to them. Spectral measurement of histamine binding to P450 in liver microsomes resolved high and intermediate affinity binding sites (Ks1 = 2.4 ± 1.6 μM; Ks2 = 90 ± 17 μM) that corresponded to microsomal binding sites (Kd1 = 1.0 ± 0.9 μM; Kd2 = 57 ± 13 μM) resolved by 3H-histamine binding; additional low affinity (Kd3 ∼ 3 mM), and probably physiologically irrelevant, sites were resolved only by 3H-histamine radioligand studies. As determined spectrally, treatment of microsomes with NADPH/carbon monoxide decreased histamine binding to P450 by about 90% and, as determined by 3H-histamine binding, abolished the high affinity sites and reduced by 85% the number of intermediate sites. Spermine competed potently for 3H-histamine binding: in microsomes, Ki = 9.8 ± 5.8 μM; in nuclei, Ki = 13.7 ± 3.1 μM; in chromatin, Ki = 46 ± 33 nM. Polyamines inhibited the P450/histamine absorbance complex with the rank order of potency: spermine > spermidine ≫ putrescine. In contrast, histamine did not compete for 3H- spermidine binding in nuclei or microsomes, suggesting that polyamines modulate histamine binding allosterically. We propose that certain P450 isozymes that modulate gene function by controlling the level of oxygenated lipids, represent at least one common intracellular target of growth-regulatory endogenous bioamines and, as shown previously, of exogenous growth-modulatory drugs including antiestrogens, antiandrogens, and certain antidepressants and antihistamines. J. Cell. Biochem. 69:233–243, 1998. © 1998 Wiley-Liss, Inc. 相似文献
19.
Yuliia N. Nekrasova Vladimir B. Sadovnikov Yury A. Zolotarev Elena V. Navolotskaya 《Journal of peptide science》2010,16(6):263-268
The synthetic peptide TPLVTLFK corresponding to the sequence 12–19 of β‐endorphin (referred to as octarphin) was found to bind to high‐affinity naloxone‐insensitive binding sites on membranes isolated from the rat brain cortex (Kd = 2.6 ± 0.2 nM ). The binding specificity study revealed that these binding sites were insensitive not only to naloxone but also to α‐endorphin, γ‐endorphin, [Met5]enkephalin, and [Leu5]enkephalin, as well. The [3H]octarphin specific binding with brain membranes was inhibited by unlabeled β‐endorphin (Ki = 2.4 ± 0.2 nM ) and a selective agonist of nonopioid β‐endorphin receptor decapeptide immunorphin SLTCLVKGFY (Ki = 2.9 ± 0.2 nM ). At the same time, unlabeled octarphin completely (by 100%) inhibited the specific binding of [3H]immunorphin with membranes (Ki = 2.8 ± 0.2 nM ). Thus, octarphin binds with a high affinity and specificity to nonopioid receptor of β‐endorphin on rat brain cortex membranes. Copyright © 2010 European Peptide Society and John Wiley & Sons, Ltd. 相似文献
20.
2-Deoxy-d -glucose (2 DG) entered synaptosomes (from rat brain) by a high-affinity, Na+-independent glucose transport system with a Km, of 0.24 mM. 3-O-methyl-glucose, D-glucose, and phloretin were competitive inhibitors of 2-DG transport with Ki's of 7 mM, 64 μM, and 0·75 μM, respectively. Insulin was without effect. 2-DG uptake was also saturable at high substrate concentrations with an apparent low affinity Km, of 75 mM, where the Kl, for glucose was 17.5 mM. We are not certain whether the rate-limiting step for the low-affinity uptake system is attributable to transport or phosphorylation. However, the high-affinity glucose transport system probably is a special property of neuronal cell membranes and could be useful in helping to distinguish separated neurons from glial cells. 相似文献