A graphic method for the determination and presentation of binding parameters in a complex system

Rosenthal, Hannah E.

doi:10.1016/0003-2697(67)90297-7

Cited by 1,496 publications

(439 citation statements)

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“…Values for ER were obtained by linear regression analysis of double reciprocal plots of the saturation data. Values for PR were obtained after applying the correction protocols of Rosenthar 35 and Feldman 38 to saturation data plotted according to the method of Scatchard. 33 Tissues were from the same baboons described in Table 1.…”

Section: Discussionmentioning

confidence: 99%

“…Two classes of aortic progesterone binding components were detected. These data were analyzed by the method of Scatchard 33 using the corrections described by Rosenthai 35 and Feld-man. 38 One class was of high affinity and limited capacity as previously described 37 and was comparable to type I estrogen receptors.…”

Section: Preparation Of Cytoplasmlc Extracts and Quantification Of Tomentioning

confidence: 99%

See 1 more Smart Citation

Gender and baboon aortic steroid hormone receptors.

Lin¹,

González²,

Carey³

et al. 1987

Arteriosclerosis

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To examine the potential of steroid hormones to serve as putative regulators of aortic cell function, we defined hormone receptor content and distribution In intact baboons. Total androgen receptor content In baboon aortic arch, thoracic arch, and abdominal aorta of young mature males was Indistinguishable from that of proestrus females. However, 30% to 40% of male aortic androgen receptors were In the nuclear fraction, whereas all aortic androgen receptors of proestrus females were In the cytoplasmlc fraction. Cytoplasmlc fraction estrogen receptor content of aortic arch and thoracic aorta of Intact males was indistinguishable from that of proestrus females. However, cytoplasmlc fraction estrogen receptor content of abdominal aorta of proestrus females was significantly greater than that of males. Nuclear fraction estrogen receptors were not detectable In either male or proestrus female baboon aortas. To assess effects of endogenous estrogen on aortic progesterone receptor content, we quantified cytoplasmlc fraction progesterone receptors and found that content of proestrus female aortic arch was not significantly different from that of males. However, cytoplasmlc fraction progesterone receptor content of thoracic and abdominal aorta of proestrus females was significantly higher than that of males. To determine whether differences In aortic receptor content or distribution were associated with changes In aortic cell function, we quantified the activity of two enzymes of glycosamlnoglycan metabolism. Aortic B-glucuronldase activity was not different In male or proestrus female baboons. Urldlne dlphosphate glucose (UDPG) dehydrogenase activity of aortic arch and thoracic aorta of proestrus female baboons was not different from that of males; however, UDPG dehydrogenase activity of abdominal aorta of proestrus females was greater than that of males. Our studies establish endogenous androgen regulation of Intracellular distribution of baboon aortic androgen receptors and imply that these receptors are physiologically functional. The higher cytoplasmlc fraction progesterone receptor content of female thoracic and abdominal aorta, as compared to that of males, suggests aortic estrogen receptors also are physiologically functional. These findings establish that baboon aortic steroid hormone receptor content and distribution Is sexually dimorphic, as Is the case for rodents. However, the essential Identity of B-glucuronldase or UDPG dehydrogenase activity In male and female baboon aortas Indicates that baboon aortic glycosamlnoglycan metabolism Is not sexually dimorphic. The fact that aspects of aortic steroid hormone receptor homeostasls and steroid hormone regulation of aortic UDPG dehydrogenase and B-glucuronldase activity significantly differ In baboons and rodents Implies complex effects of steroid hormones which may be speciesspecific. (Arteriosclerosis 7:248-255, May/June 1987) I t is well established that men have a higher incidence of coronary heart disease (CHD) than age-matched females; however, the underlyi...

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Section: Discussionmentioning

confidence: 99%

Section: Preparation Of Cytoplasmlc Extracts and Quantification Of Tomentioning

confidence: 99%

Gender and baboon aortic steroid hormone receptors.

Lin¹,

González²,

Carey³

et al. 1987

Arteriosclerosis

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“…If the polymer has more than one group of binding sites, a curved line is obtained in the Scatchard plot rather than a straight line 35 . In the case of one ligand (metergoline) and two types of binding sites (one type of site with high and one with low selectivity) two straight lines would be generated with different slopes according to the Scatchard equation.…”

Section: Scatchard Analysismentioning

confidence: 99%

“…Therefore, the values of and should be considered as apparent values. They can be used for comparative purpose but cannot be regarded as absolute values considering the assumptions made in the Scatchard approach 32,31,35 . However, they are a good indicator for the presence of different binding sites on the MIP as a curved experimental slope is observed.…”

Section: Scatchard Analysismentioning

confidence: 99%

Development of Suspension Polymerized Molecularly Imprinted Beads with Metergoline as Template and Application in a Solid-Phase Extraction Procedure toward Ergot Alkaloids

Lenain

Mavungu²,

Dubruel³

et al. 2012

Anal. Chem.

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The first successfully developed molecularly imprinted polymer toward six ergot alkaloids and their respective epimers is described. A new imprinting molecule, metergoline, was used as template analogue in the production of suspension polymerized beads. These spherical particles functioned as selective sorbent in a solid-phase extraction column. The application of this column in the cleanup of barley samples prior to liquid chromatography coupled with tandem mass spectrometry allowed simple and cost-efficient sample preparation. The performance of the imprinted polymer and a non-imprinted control polymer was evaluated. This includes determination of the recovery values and the matrix effect of each of the 12 tested ergot alkaloids as well as a crossreactivity study with 25 common mycotoxins. The binding isotherms were obtained for metergoline, thus allowing comparison with other (imprinted) sorbents. A comparison between bulk and suspension polymerization is provided to determine the appropriate production technique. E rgot alkaloids are mycotoxins produced by Claviceps species, specifically Claviceps purpurea. These fungi can be found on cereals and produce sclerotia, which vary in composition and concentration of ergot alkaloids. The sclerotia are harvested together with the grains and only up to 80% of the kernels can be removed with the mechanical separation techniques currently available. Especially in periods of drought these techniques become unreliable as smaller sclerotia, similar in size to the grains, are produced. Generally, sclerotia are also broken during transport, allowing them to enter the food chain more easily.These ergot alkaloids possess toxicity due to interactions with α-adrenergic, serotinergic, and dopaminergic receptors. 1−3 Excess intake by humans can cause nausea, convulsions, hallucinations, and vasoconstriction and can even lead to gangrenous symptoms and abortion. Animals are infected through consumption of contaminated feed. This is a widespread phenomenon in livestock leading to adverse effects ranging from weight loss to death of infected animals. [4][5][6]2,7 The European Food Safety Authority (EFSA) stated in 2005 that the degree of variability in ergot alkaloid pattern in relation to fungal species, host plant, and geographical distribution is not known. 5 Six ergot alkaloidsergometrine, ergosine, ergotamine, ergocristine, ergocryptine, and ergocornine which are considered the most important ones according to EFSA, are depicted in Figure 1. An isomerization from the biologically active R-form to the biologically inactive S-form occurs at the C 8 stereogenic center of the tetracyclic ergoline structure in a reversible epimerization reaction. 8,9,6 The suffix "-ine" is used in the nomenclature if the side chain is present in the R-form at the C 8 stereogenic center, for example, ergotamine. The side chain is present in the S-orientation in the corresponding epimer which is expressed by the suffix "-inine", for example, ergotaminine. This reaction is promoted in humid, aqueous,...

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“…Specific binding was determined as described above. Results were analysed by the method of Rosenthal (23).…”

Section: P R In Cerebellum and Hypothalamus Throughout The Oestrous Cmentioning

confidence: 99%

Oestrogen‐Insensitive Progestin Receptors in the Central Nervous System: Physicochemical and Immunoreactive Characteristics

Cerbón¹,

Martineíz²,

Pérez‐Palacios³

1989

J Neuroendocrinology

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The intracellular effects of progesterone in the central nervous system are exerted via two distinct receptors: The classical oestrogen-regulated progestin receptor and the non-oestrogen-inducible progestin receptor. To assess whether the oestrogeninsensitive receptor is related to the oestrogen-dependent receptor or whether it is a different binding macromolecule, its physicochemical and immunoreactive characteristics (immunoprecipitation with polyclonal anti-uterine progesterone receptor antisera) were studied in neural tissues of the female rat. The results disclosed that the dissociation constant (Kdr 0.4-0.5 x M), stereospecificity and sedimentation coefficient (7-8 S ) in linear sucrose gradients of the oestrogen-insensitive progestin receptor were identical to those reported for the oestrogen-regulated progestin receptor, although its saturation binding capacity was significantly lower. Results of in vifro nuclear acceptor assays revealed that the progestin receptor complexes from cerebellum and cerebral cortex, were able to specifically bind to cell nuclei preparations in a fashion similar to that observed with the uterine progestin receptor, although to a lesser extent. Interestingly, a similar nuclear uptake of receptor complexes was noticed when standardized cerebellum and uterus cytosol preparations with a similar receptor content were used. The anti-uterine progesterone receptor immunoglobulins used in the immunoprecipitation studies were able to specifically recognize the progestin receptor populations of the anterior pituitary and hypothalamus (oestrogen-regulated receptors) as well as those of the cerebellum and cerebral cortex (oestrogen-insensitive receptors). The results presented show that both the oestrogen-sensitive and the oestrogeninsensitive cytosol progestin receptors in brain bind the same progesterone-like molecules used as radioligands and also react with the same antibody when tested in an immunoprecipitation radioassay. The striking similarities found in binding kinetics, physicochemical characteristics and immunoreactive behaviour in the two progestin receptors studied demonstrated that both macromolecules belong to the same family of proteins in spite of their different sensitivity to oestrogens. The overall data seem to suggest a common origin of the two progestin receptor populations in brain but with different mechanisms of hormonal regulation.

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A graphic method for the determination and presentation of binding parameters in a complex system

Cited by 1,496 publications

References 2 publications

Gender and baboon aortic steroid hormone receptors.

Gender and baboon aortic steroid hormone receptors.

Development of Suspension Polymerized Molecularly Imprinted Beads with Metergoline as Template and Application in a Solid-Phase Extraction Procedure toward Ergot Alkaloids

Oestrogen‐Insensitive Progestin Receptors in the Central Nervous System: Physicochemical and Immunoreactive Characteristics

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