The kinetics of estrogen binding to rat alpha-fetoprotein

Fluid obtained from rat fetuses was utilized to characterize the affinity, number of binding sites, and the association and dissociation rate kinetics of the binding of estradiol and estrone to AFP. Statistical analysis demonstrated no differences when the values for the AFP-estradiol interaction were compared with those obtained for the ATP-estrone interaction. These data demonstrate that rat AFP specifically binds estradiol and estrone with a high capacity, high affinity, and similar binding kinetics.     

Binding of (125I) triiodothyronine to human peripheral leukocytes and its internalization

Summary Ultrastructural autoradiography showed high specific binding of (¹²⁵I) triiodothyronine, as confirmed by a competition test, to plasma membranes, nuclei and mitochondria of human peripheral leukocytes. A high level of binding was also noted on the granulocytes' granules, especially in eosinophils.     

High affinity binding of 5-hydroxytryptamine to some membrane preparations from cattle brain. Relationship to lysergic acid diethylamide (LSD)]

5 hydroxytryptamine binds to crude brain membrane preparations with two different affinities (KD = 1 to 2 X 10(-9) M for the highest, 1 to 2 X 10(-8) M for the lowest). LSD also binds with two affinities (KD = 3 to 4 X 10(-9) M and KD = 2 to 3 X 10(-8) M). Subcellular distribution of these sites shows that binding involves the two binding affinities in microsomal membranes but solely the high affinity binding sites are present in purified synaptosomal membranes. High affinity sites for 5 HT and for LSD are different as no direct competitive inhibition is observed in that case. On microsomal membranes, direct relationship occurs between low affinity binding for 5 HT and high affinity binding for LSD.     

The kinetics of estrogen binding to rat α-fetoprotein

Summary Fluid obtained from rat fetuses was utilized to characterize the affinity, number of binding sites, and the association and dissociation rate kinetics of the binding of estradiol and estrone to AFP. Statistical analysis demonstrated no differences when the values for the AFP-estradiol interaction were compared with those obtained for the AFP-estrone interaction. These data demonstrate that rat AFP specifically binds estradiol and estrone with a high capacity, high affinity, and similar binding kinetics.This work was supported by NSF grant PCM-8109847 and by a Grant-in-Aid of Research from Sigma Xi, The Scientific Research Society.     

Binding capacity of the testosterone-binding plasma protein in 2 wild mammals with cyclic testicular activity, the fox and badger

The binding capacity of the plasma testosterone binding globulin (TeBG) as determined by the equilibrium dialysis method, in fox and badger, was shown to wary in relation to the seasonal endocrine activity of the testis. During the winter breeding period, the TeBG binding capacity was low in the fox and high in the badger, whereas an opposite situation prevailed during the sexual quiescence phase. Independent of those seasonal variations, the binding capacity of TeBG was always lower in fox than in badger.     

Effect of phosphorylation of myosin light chains on interaction of heavy meromyosin with regulated F-actin in ghost fibers

The binding of phosphorylated heavy meromyosin to regulated F-actin in ghost fibers at high Ca2+ concentration increases, and at low Ca2+ concentration decreases, the anisotropy of intrinsic tryptophan fluorescence of F-actin. The effect is opposite to the effect of the binding of dephosphorylated heavy meromyosin.     

Binding of 11-cis retinaldehyde to the partially purified cellular retinaldehyde binding protein from bovine retinal pigment epithelium

11-cis retinaldehyde binding analysis was performed on a bovine retinal pigment epithelium preparation of cellular retinaldehyde binding protein (CRALBP), whose purity degree was estimated as 75%. Equilibrium binding studies were carried out measuring the replacement of tritium-labeled with unlabeled 11-cis retinaldehyde at 25 degrees C. Analysis of the experimental data both by a direct curve-fitting procedure utilizing a non linear least square regression analysis and by a conventional Scatchard plot revealed a single non-interacting binding site with an apparent equilibrium constant of 0.9 X 10(-7) M. A binding stoichiometry of approximately 1 mol of 11-cis retinaldehyde/mol of binding protein can be calculated from the experimental data. Competition studies carried out in the presence of unlabeled 'trans' and 'cis' isomers of vitamin A derivatives confirm the high degree of specificity of the 11-cis retinaldehyde binding.     

Identification of calcium antagonist receptor binding sites using (3H)nitrendipine in bovine tracheal smooth muscle membranes

(3H)Nitrendipine binding to the bovine tracheal muscle membrane at 25 degrees C was rapid, saturable (Bmax = 14.8 +/- 3.9 fmol/mg protein) and of high affinity (Kd = 0.15 +/- 0.04 nM). The rank order of Ca2+ antagonists competing for airway (3H)nitrendipine binding was nitrendipine not equal to nisoldipine not equal to nifedipine much greater than verapamil. Cromolyn, however, neither inhibited nor increased the binding.     

Androgen receptors in the cytosol of normal mucosa and of pharyngo-laryngeal epitheliomas in humans]

Specific binding for 5 alpha dihydrotestosterone has been characterized in cytosol fraction from human pharyngeal and laryngeal mucosae. The high binding affinity of the protein receptor (Kd 2 x 10(-10)M) and its low capacity are similar in both sexes, and similar in both normal mucosa and epithelioma.     

Visualization of opiate receptors in the locus coeruleus of rats: high resolution radioautography after administration of a tritiated met-enkephalin analog]

Opiate receptor sites were visualized by high resolution radioautography in the locus coeruleus of the Rat following intraventricular injection of the met-enkephalin analogue FK 33-824 tritiated with a high specific activity (3H-FK). After administration of a tracer dose of 3H-FK, more than 75% of the radioactivity detected in the locus coeruleus is specifically bound to opiate receptor sites. These are distributed both between and inside neuronal perikarya. After administration of 3H-FK at high concentration, electron microscope radioautography shows the presence of specific opiate binding sites at the level of axo-somatic and axo-dendritic synaptic junctions. These synaptic binding sites could correspond to receptor sites for endogenous enkephalins.     

3[H]-Sulpiride labels mesolimbic non-dopaminergic sites that bind antidepressant drugs

3[H]-(-)-Sulpiride and 3[H]-spiperone binding was compared in rat amygdala, nucleus accumbens and striatum, using (+/-)-sulpiride to define specific binding. 3[H]-(-)-Sulpiride bound to twice as many sites in amygdala and nucleus accumbens as 3[H]-spiperone. 3[H]-(-)-Sulpiride binding was directed to these additional sites by using 1 microM spiperone to mask dopaminergic binding. The binding of 3[H]-(-)-sulpiride to these sites was high affinity, reversible, Na+-dependent, but not stereospecific. Metoclopramide, tiapride and antidepressant medications, but not other neuroleptics, ADTN, or serotonin displaced 3[H]-(-)-sulpiride binding to these sites. These data suggest that 3[H]-(-)-sulpiride labels mesolimbic sites other than dopamine receptors which may mediate antidepressant effects.     

Butanol extracts from myelin fragments. IV. Some interactions between 5-hydroxytryptamine and other neurotransmitters binding

Summary To examine the interaction between 5-HT and other neurotransmitters binding to the butanol extracts from myelin, double labelling experiments were done. The binding peaks of C¹⁴. ACh and NA were clearly different from that of H³. 5-HT. At 5×10^–7 M, binding of 5-HT, ACh, NA, GABA and DA was 62.7, 2.3, 7.0, 5.8 and 1.9 nmoles/mg protein, respectively. These results suggest that the 5-HT binding components of myelin butanol extracts may have high selectivity and specificity.     

Effect of phosphorylation of myosin light chains on interaction of heavy meromyosin with regulated F-actin in ghost fibers

Summary The binding of phosphorylated heavy meromyosin to regulated F-actin in ghost fibers at high Ca²⁺ concentration increases, and at low Ca²⁺ concentration decreases, the anisotropy of intrinsic tryptophan fluorescence of F-actin. The effect is opposite to the effect of the binding of dephosphorylated heavy meromyosin.     

Tissue specificity of testosterone fixation: demonstrated by ultrastructural immunocytochemistry after cryoultramicrotomy]

Testosterone can be detected by immunocytochemistry on ultrathin slices obtained by cryoultramicrotomy. Testosterone detected by this method is likely bound on a binding site having a high affinity. In order to study the tissue specificity of this binding, testosterone was tested in Rat pituitary gland, liver and adrenal glands. Immunoreactive testosterone was detected in the gonadotropic cells of the pituitary gland, in the hepatocytes. Testosterone was not detected by immunocytochemistry in the pituitary cells other than the gonadotropic cells and in the cells of the medulla of adrenal glands. These data testify in favour of tissue specificity of the testosterone binding.     

Extraction and purification of brain glycoprotein NSA3 by binding with hyaluronic acid

Brain glycoprotein NSA3 was found to bind to immobilised hyaluronic acid. The binding was reversible and gave pure antigen (99%) in high yields (80%). The binding was suppressed by incubating the affinosorbent with hyaluronidase. It was not suppressed by trypsin. The presence of glycosaminoglycans other than hyaluronic acid in the sample did not inhibit the binding. This property is relevant to those already known for the brain glycoprotein NSA3, and to its localisation at the nodes of Ranvier. We propose to coin the name hyaluronectin for the brain glycoprotein NSA3.     

Behaviour of intrinsically disordered proteins in protein–protein complexes with an emphasis on fuzziness

Intrinsically disordered proteins (IDPs) do not, by themselves, fold into a compact globular structure. They are extremely dynamic and flexible, and are typically involved in signalling and transduction of information through binding to other macromolecules. The reason for their existence may lie in their malleability, which enables them to bind several different partners with high specificity. In addition, their interactions with other macromolecules can be regulated by a variable amount of chemically diverse post-translational modifications. Four kinetically and energetically different types of complexes between an IDP and another macromolecule are reviewed: (1) simple two-state binding involving a single binding site, (2) avidity, (3) allovalency and (4) fuzzy binding; the last three involving more than one site. Finally, a qualitative definition of fuzzy binding is suggested, examples are provided, and its distinction to allovalency and avidity is highlighted and discussed.     

Presence of specific binding sites for [3H]sarcophytol-A in cultured human skin fibroblasts

The presence of specific binding sites for [3H]sarcophytol-A in human skin fibroblasts was examined using biochemical and morphological methods. The displacement studies clearly revealed that high (KD = 31.0 nM) and low (KD = 6.05 microM) affinity sites were present in the intact cells. Moreover, autoradiographic studies using light microscopy revealed that the specific binding sites may exist in both the cytoplasm and the nuclei.     

Immunochemical similarities among the hemolymph juvenile hormone binding proteins of moths

The hemolymph from various species of moths was analyzed for cross-reactivity with a panel of six monoclonal antibodies made against the hemolymph juvenile hormone binding protein ofManduca sexta. With the exception of one antibody, the immunoreactivity was limited to the sphingid family. One monoclonal antibody cross-reacted with a number of lepidopteran species; however, families such as Noctuidae and Pyralidae, known to have high affinity, low molecular weight juvenile hormone binding proteins, did not cross-react. Immunological cross-reactivity withManduca sexta juvenile hormone binding protein in several primitive moth families supports the current model of phylogenetic relationships in the order Lepidoptera.     

Opiate receptor sites in neuronal and glial cell lines in culture]

A stereospecific saturable high affinity binding of 3H-naloxone has been found in 3 glial and 2 neuronal cell lines. Kinetic study of binding seems to indicate only one class of receptor sites in the 5 cell lines. Acute exposure to morphine (1 X 10(-5)M) concurrent with PGE1-induced stimulation of adenylate cyclase did not result in a decrease of the cAMP level in any cell line tested.     

Observations on salicyl hydroxamic acid,an experimental trypanocide

Summary The inferior in vivo efficacy of salicyl hydroxamic acid against trypanosomiasis may be explained by its short half-life, high degree of protein binding and low tissue levels.