Insulin interactions with its receptors: Experimental evidence for negative cooperativity

Meyts, Pierre De; Roth, Jesse; Neville, David M.; Gavin, James R.; Lesniak, Maxine A.

doi:10.1016/s0006-291x(73)80072-5

Cited by 717 publications

(365 citation statements)

References 21 publications

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“…Experimental support for this suggestion is afforded by the observation that the rate of dissociation of radioactively labelled insulin is increased in the presence of an excess of native insulin (De Meyts et al, 1973, 1976. We have confirmed these observations in our preparation of rat liver plasma membranes, but, significantly, bovine plasma membranes do not show this effect.…”

Section: Discussionsupporting

confidence: 80%

“…It has been suggested that this observation in other receptor preparations may be due to site-site interactions of the negative co-operative type (De Meyts et al, 1973, 1976. Experimental support for this suggestion is afforded by the observation that the rate of dissociation of radioactively labelled insulin is increased in the presence of an excess of native insulin (De Meyts et al, 1973, 1976.…”

Section: Discussionmentioning

confidence: 99%

“…For studies with insulin analogues, plasma membranes were incubated in duplicate with a fixed amount of 1251-labelled insulin (0.25nM) and vari- Dissociation ofbound insulin Dissociation of bound insulin from the membranes was studied as described by De Meyts et al (1973, 1976.…”

Section: Insulin Bindingmentioning

confidence: 99%

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Binding of insulin to bovine liver plasma membrane. Use of insulin analogues modified at the A1 residue

Rösen

Simón

Reinauer

et al. 1980

Biochemical Journal

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Bovine liver plasma membranes [Rosen, Ehrich, Junger, Bubenzer & Kuhn (1979) Biochim. Biophys. Acta 587, 593-6051 show similar insulin-binding characteristics, as evaluated by Scatchard analysis, to those of membrane systems from other species. However, the dissociation rate of bound insulin cannot be accelerated by the addition of insulin, in contrast with membranes isolated from rat liver. The dissociation rate is strongly dependent on the pH. Although dependent on temperature, the total capacity of binding sites is minimally changed, but the number of high-affinity sites is increased 2-3-fold, by lowering the incubation temperature. These data might be interpreted by assuming a single population of receptors whose distribution between different affinity states depends on temperature. In competition studies, most of the modified insulins examined show a close correlation between binding, determined i'n plasma membranes from bovine liver, and biological activity, measured in adipocytes. The hypothesis that a positive charge on the Al residue may be favourable for binding is supported by experiments with an isosteric pair of insulins modified at this residue ([carbamoylGlyAl]-and [amidino-GlyAllinsulin) and with modified insulins carrying one or more positive charges on the Al residue ([Arg-GlyAl1-, [Arg-Arg-GlyAl1-, [Arg-ArgArg-GlyAl1-and [Lys-Arg-GlyAl]insulin). The latter insulin derivatives show a higher binding activity for plasma membranes from bovine, porcine and rat liver than expected from their biological activities in adipocytes.

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Section: Discussionsupporting

confidence: 80%

Section: Discussionmentioning

confidence: 99%

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Binding of insulin to bovine liver plasma membrane. Use of insulin analogues modified at the A1 residue

Rösen

Simón

Reinauer

et al. 1980

Biochemical Journal

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“…A salient manifestation of this mechanism is the ability of unlabelled competitive ligands to accelerate the dissociation of a prebound radioligand both in ex vivo [70][71][72][73] and in vitro washout experiments [15,74,75]. It is noteworthy that 'rebinding' occurs at the submicroscopic level [76] and, hence, that it should not be confounded by earlier 'macroscopic' definitions such as the restoration of a new equilibrium between targets and dissociated radioligand molecules [77,78].Research in different life science disciplines supports the perception that the anatomical complexity of intact tissues encourages rebinding to a much higher extent than the simple presence of the target-bearing membrane alone. Most compellingly, ex vivo experiments reported by Sadée et al [71] revealed that, whereas the D 1 dopamine receptor antagonist [ 3 H]-SCH 23390 experienced sizable rebinding in 'still living' striatal slices of the rat, this was no longer the case after tissue homogenization.…”

mentioning

confidence: 99%

Cell membranes… and how long drugs may exert beneficial pharmacological activity in vivo

Vauquelin

2016

Brit J Clinical Pharma

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The time course of the beneficial pharmacological effect of a drug has long been considered to depend merely on the temporal fluctuation of its free concentration. Only in the last decade has it become widely accepted that target-binding kinetics can also affect in vivo pharmacological activity. Although current reviews still essentially focus on genuine dissociation rates, evidence is accumulating that additional micro-pharmacokinetic (PK) and -pharmacodynamic (PD) mechanisms, in which the cell membrane plays a central role, may also increase the residence time of a drug on its target. The present review provides a compilation of otherwise widely dispersed information on this topic. The cell membrane can intervene in drug binding via the following three major mechanisms: (i) by acting as a sink/repository for the drug; (ii) by modulating the conformation of the drug and even by participating in the binding process; and (iii) by facilitating the approach (and rebinding) of the drug to the target. To highlight these mechanisms, we focus on drugs that are currently used in clinical therapy, such as the antihypertensive angiotensin II type 1 receptor antagonist candesartan, the atypical antipsychotic agent clozapine and the bronchodilator salmeterol. Although the role of cell membranes in PK-PD modelling is gaining increasing interest, many issues remain unresolved. It is likely that novel biophysical and computational approaches will provide improved insights in the near future. IntroductionThe pharmacokinetic (PK) and pharmacodynamic (PD) properties of a drug are determinants of its efficacy and the duration of its clinical action [1]. PK and PD have long been considered to be separate disciplines, and the time course of the pharmacological effect of a drug has essentially been considered in terms of the temporal fluctuation of its free concentration [2]. In accordance with this principle, the frequently observed time lag between the concentration of a drug in the systemic circulation and its effect was initially attributed to slow equilibration between the plasma and a hypothetical target-surrounding 'effect compartment' [3]. By contrast, preclinical screening studies traditionally aim to optimize drug candidates in terms of only their efficacy and potency (i.e. PD properties). Inherent to this practice is the proposal that drug-target interactions are adequately described by equilibrium equations and, hence, that association and dissociation rates play only subordinate roles. In addition to a few noteworthy exceptions [4,5], it is only in the last decade that it has become fashionable to include binding kinetics as an additional criterion for clinical efficacy. This insight was largely sparked by the seminal articles by Swinney [6] and Copeland et al. [7]. The numerous review articles that have been published subsequently have focused mainly on long residence times. This property is now recognized to play a key role with respect to the clinical British Journal of Clinical Pharmacology Br J Clin Pharmacol (2016...

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“…However, the concave upward curve shown in figure 7 could also be interpreted in terms of negative cooperativity (28,29). One has to realize that in the presence of cooperative mechanisms the values of the dissociation constant and the number of binding sites derived from a Scatchard analysis do not have their usual precise physico-chemical meaning.…”

Section: Discussionmentioning

confidence: 99%

Preparation of Parenchymal and Non-Parenchymal Cells from Adult Human Liver — Morphological and Biochemical Characteristics

Bojar¹,

Basler²,

Fuchs³

et al. 1976

Clinical Chemistry and Laboratory Medicine

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Summary: By perfusion of the isolated human liver with collagenase and hyaluronidase a mixed suspension of various cell types was obtained. Pure parenchymal cells were prepared by differential centrifugation, pure non-parenchymal cells by the use of pronase and subsequent isopycnic centrifugation on metrizamide gradients (50-300 g/1). About 90% of the parenchymal and non-parenchymal cells were viable as judged by tiypan blue staining. Non-parenchymal cells were not capable of gluconeogenesis but utilized glucose at high rates. Parenchymal cells retained their ability to form glucose and to accumulate glycogen from fructose > lactate/pyruvate > alanine. Studies on binding of 125 Ilabelled insulin by isolated parenchymal cells were performed at 30 °C. The binding data may fit a model with a minimum of two classes of binding sites: (a) high affinity -low capacity sites (Kd ~~ 6.6 nmol/1, capacity ~ 16000 insulin molecules per cell) and (b) low affinity -high capacity sites (K d ~ 0.37 / , capacity ~ 646 000 molecules per cell).

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Insulin interactions with its receptors: Experimental evidence for negative cooperativity

Cited by 717 publications

References 21 publications

Binding of insulin to bovine liver plasma membrane. Use of insulin analogues modified at the A1 residue

Binding of insulin to bovine liver plasma membrane. Use of insulin analogues modified at the A1 residue

Cell membranes… and how long drugs may exert beneficial pharmacological activity in vivo

Preparation of Parenchymal and Non-Parenchymal Cells from Adult Human Liver — Morphological and Biochemical Characteristics

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