scholarly journals Subcutaneously Delivered Nano-Insulin Drug as Label Free Nanotheranostic

2021 ◽  
Author(s):  
Rıdvan Say ◽  
Almila Şenat ◽  
Özlem Biçen Ünlüer ◽  
Fahrettin Akyüz ◽  
Arzu Ersöz
Keyword(s):  
Liquid Chromatography ◽  
Insulin Receptor ◽  
Rat Liver ◽  
Insulin Receptors ◽  
Insulin Analog ◽  
Second Step ◽  
Label Free ◽  
Column Material ◽  
Reflectometric Interference Spectroscopy ◽  
The Relationship

Abstract In this study, it has aimed to understand the relationship between purified insulin and insulin receptor, also nanoinsulin and insulin receptors. The insulin receptor has separated from rat liver using a cryogel column material that is photosensitively insulin cross-linked in the Fast Protein Liquid Chromatography (FPLC) system based on the affinity between insulin and insulin receptor. In the second step, an isolated insulin receptor has used to synthesize insulin receptor cross-linked cryogels for purifying insulin from rats. Subcutaneously delivered nano-insulin drug has prepared from the purified insulin using AmiNoAcid (monomer) Decorated and Light Underpinning Conjugation Approach (ANADOLUCA) method. Lastly, Reflectometric Interference Spectroscopy (RIfS) study has performed to understand the interaction between purified insulin receptor and purified insulin, commercial insulin analog, and nano insulin. These studies have demonstrated that nano-insulin drugs can be effectively used as a theranostic platform to monitor affinity and blocking interactions of nanoprotein drug and its receptor.

scholarly journals Tyrosine phosphorylation of two cytosolic proteins of 50 kDa and 35 kDa in rat liver by insulin-receptor kinase in vitro

1987 ◽  
Vol 248 (1) ◽  
pp. 27-33 ◽  
Author(s):  
Y C Kwok ◽  
C C Yip
Keyword(s):  
Insulin Receptor ◽  
Rat Liver ◽  
Growth Factor Receptor ◽  
Insulin Receptors ◽  
Artificial Substrates ◽  
Receptor Kinase ◽  
Insulin Receptor Tyrosine Kinase ◽  
Cytosolic Proteins ◽  
Insulin Receptor Kinase

Insulin-receptor tyrosine kinase can phosphorylate a variety of artificial substrates in vitro. Its physiological substrate(s), however, remains unknown. In the present study, we show that immobilized insulin receptors phosphorylate tyrosine residues of two cytosolic proteins of 50 kDa and 35 kDa in rat liver. Phosphorylation of these two proteins required Mn2+- or Mg2+-ATP as the phosphate donor. Phosphorylation was time- and temperature-dependent. Furthermore, the rate of phosphorylation of the two proteins was related to the autophosphorylated state of the insulin receptor. The pI of the phosphorylated 50 kDa and 35 kDa proteins was 5.4 and 5.6 respectively. These proteins were present in low abundance. They were not related to each other, nor to the insulin receptor, as demonstrated by in-gel proteolytic digestion and by immunoprecipitation using antibodies produced against them. They were specific substrates for the insulin receptor kinase, since they were not phosphorylated by epidermal-growth-factor-receptor kinase. These observations suggest that the 50 kDa and 35 kDa cytosolic proteins may be endogenous substrates for the insulin-receptor kinase.

scholarly journals Analysis of insulin receptor phosphorylation sites in intact rat liver cells by two-dimensional phosphopeptide mapping. Predominance of the tris-phosphorylated form of the kinase domain after stimulation by insulin

1991 ◽  
Vol 275 (1) ◽  
pp. 15-21 ◽  
Author(s):  
T Issad ◽  
J M Tavaré ◽  
R M Denton
Keyword(s):  
Insulin Receptor ◽  
Rat Liver ◽  
Insulin Receptors ◽  
Kinase Domain ◽  
Two Dimensional ◽  
Tyrosine Residues ◽  
C Terminus ◽  
Rat Liver Cells ◽  
Wheat Germ Lectin ◽  
Dimensional Mapping

1. Insulin receptors were partially purified from rat liver by chromatography on wheat-germ-lectin-Sepharose. Incubation with [gamma-32P]ATP in the presence of insulin resulted in increased phosphorylation of the beta-subunit on both tyrosine and serine residues. Two-dimensional mapping of tryptic peptides showed that, in agreement with previous studies using preparations of receptors from other sources, the tyrosine residues involved were the three tyrosines in the kinase domain (corresponding to tyrosines 1158, 1162 and 1163 of the human receptor) plus two tyrosines close to the C-terminus (corresponding to tyrosines 1328 and 1334). 2. The effects of insulin on the phosphorylation of receptors within intact rat liver cells were determined by incubating cells in the presence of [32P]Pi for 50 min and then with or without insulin for a further 10 min. The labelled receptors were then rapidly isolated by sequential use of wheat-germ-lectin-Sepharose chromatography and immuno-isolation using a monoclonal antibody to the C-terminal end of the beta-subunit. 3. Insulin was found to increase overall phosphorylation of the receptor nearly 3-fold. Two-dimensional mapping was then carried out in combination with phosphoamino acid analysis. This revealed that the pattern of phosphorylation of the receptors in cells incubated in the absence and presence of insulin exhibited a number of marked differences from that observed in previous studies on intact cells, which had been restricted to cells expressing very high levels of insulin receptors such as certain hepatoma-derived cells or cells transfected with insulin receptor cDNA. The differences in the effects of insulin included a larger increase in the proportion of receptors being phosphorylated on the three tyrosine residues of the kinase domain, no apparent phosphorylation of the two tyrosine residues close to the C-terminus and no increase in either threonine or overall serine phosphorylation. 4. The receptors appeared to be phosphorylated on a number of different serine residues in cells incubated in the absence of insulin. Evidence for both increases and decreases in the phosphorylation of specific serine residues on addition of insulin was obtained. 5. It is concluded that care should be taken when extrapolating findings on the phosphorylation of the insulin receptor within cultured cells to more physiological situations.

Antiserum to liver insulin receptor hyperpolarizes rat muscle

1983 ◽  
Vol 244 (1) ◽  
pp. C58-C60 ◽  
Author(s):  
K. Zierler ◽  
E. M. Rogus
Keyword(s):  
Skeletal Muscle ◽  
Insulin Receptor ◽  
Rat Liver ◽  
Rabbit Antiserum ◽  
Insulin Receptors ◽  
Metabolic Effects ◽  
Metabolic Responses ◽  
Maximum Effect ◽  
Half Maximum ◽  
Rat Muscle

Antibodies to insulin receptors have been reported to have some insulinlike metabolic effects. If insulin-induced electrical hyperpolarization of skeletal muscle is part of the transduction chain between receptor and certain metabolic responses, then receptor antiserum should hyperpolarize. The Jacobs antiserum (rabbit antiserum against rat liver insulin receptor) hyperpolarized rat caudofemoralis muscle. Maximum effect, about 4.5 mV, occurred at 1:10,000 dilution, half maximum at about 1:40,000. Maximum effect of antiserum was only as great as half maximum hyperpolarization by insulin on this muscle. 2-Deoxyglucose uptake was also stimulated by antiserum but required greater concentration than for hyperpolarization, and the stimulation was only by about one-third the maximum effect of insulin.

scholarly journals The endogenous functional turkey erythrocyte and rat liver insulin receptor is an α2β2 heterotetrameric complex

1990 ◽  
Vol 271 (1) ◽  
pp. 99-105 ◽  
Author(s):  
J L Treadway ◽  
B D Morrison ◽  
J A Wemmie ◽  
I Frias ◽  
T O'Hare ◽  
...  
Keyword(s):  
Insulin Receptor ◽  
Rat Liver ◽  
Human Placenta ◽  
Alkylating Agents ◽  
Insulin Receptors ◽  
Cross Linking ◽  
Protein Kinase Activity ◽  
Liver Membranes ◽  
Alpha Beta ◽  
Beta 2

Previous studies have indicated that turkey erythrocyte and rat liver membranes contain endogenous alpha beta heterodimeric insulin receptors in addition to the disulphide-linked alpha 2 beta 2 heterotetrameric complexes characteristic of most cell types. We utilized 125I-insulin affinity cross-linking to examine the structural properties of insulin receptors from rat liver and turkey erythrocyte membranes prepared in the absence and presence of sulphydryl alkylating agents. Rat liver membranes prepared in the absence of sulphydryl alkylating agents displayed specific labelling of Mr 400,000 and 200,000 bands, corresponding to the alpha 2 beta 2 heterotetrameric and alpha beta heterodimeric insulin receptor complexes respectively. In contrast, affinity cross-linking of membranes prepared with iodoacetamide (IAN) or N-ethylmaleimide identified predominantly the alpha 2 beta 2 heterotetrameric insulin receptor complex. Similarly, affinity cross-linking and solubilization of intact turkey erythrocytes in the presence of IAN resulted in exclusive labelling of the alpha 2 beta 2 heterotetrameric insulin receptor complex, whereas in the absence of IAN both alpha 2 beta 2 and alpha beta species were observed. Turkey erythrocyte alpha 2 beta 2 heterotetrameric insulin receptors from IAN-protected membranes displayed a 3-4-fold stimulation of beta subunit autophosphorylation and substrate phosphorylation by insulin, equivalent to that observed in intact human placenta insulin receptors. Turkey erythrocyte alpha beta heterodimeric insulin receptors, prepared by defined pH/dithiothreitol treatment of IAN-protected membranes, were also fully competent in insulin-stimulated protein kinase activity compared with alpha beta heterodimeric human placenta receptors. In contrast, endogenous turkey erythrocyte alpha beta heterodimeric insulin receptors displayed basal protein kinase activity which was insulin-insensitive. These data indicate that native turkey erythrocyte and rat liver insulin receptors are structurally and functionally similar to alpha 2 beta 2 heterotetrameric human placenta insulin receptors. The alpha beta heterodimeric insulin receptors previously identified in these tissues most likely resulted from disulphide bond reduction and denaturation of the alpha 2 beta 2 holoreceptor complexes during membrane preparation.

Insulin and insulin receptor uptake into rat liver. Chloroquine action on receptor recycling

Diabetes ◽  
1985 ◽  
Vol 34 (10) ◽  
pp. 1025-1030 ◽  
Author(s):  
M. N. Khan ◽  
S. Savoie ◽  
R. J. Khan ◽  
J. J. Bergeron ◽  
B. I. Posner
Keyword(s):  
Insulin Receptor ◽  
Rat Liver ◽  
Receptor Recycling

scholarly journals Internalization and Activation of the Rat Liver Insulin Receptor Kinase in vivo

1989 ◽  
Vol 264 (22) ◽  
pp. 12931-12940 ◽  
Author(s):  
M N Khan ◽  
G Baquiran ◽  
C Brule ◽  
J Burgess ◽  
B Foster ◽  
...  
Keyword(s):  
Insulin Receptor ◽  
Rat Liver ◽  
Receptor Kinase ◽  
Insulin Receptor Kinase

scholarly journals Ligand-induced translocation of insulin receptors in intact rat liver.

1982 ◽  
Vol 257 (18) ◽  
pp. 10852-10860 ◽  
Author(s):  
B Desbuquois ◽  
S Lopez ◽  
H Burlet
Keyword(s):  
Rat Liver ◽  
Insulin Receptors

Komponisten, Interpreten und Editoren debattieren über Notation

2019 ◽  
Vol 33 (1) ◽  
pp. 64-81
Author(s):  
Dörte Schmidt
Keyword(s):  
New York ◽  
International Association ◽  
Contemporary Music ◽  
Second Step ◽  
John Cage ◽  
Musical Structure ◽  
New Developments ◽  
The Relationship

Abstract The article discusses how new developments in the notation of contemporary music were negotiated within the framework of the Darmstadt Summer Courses and which interests and actors played a role in this. The first part examines the publications and publication projects that emerged in the context of the Notation conference in 1964. The focus is on the interests of institutions such as the International Music Council and the International Association of Music Libraries, in whose name the New York publisher Kurt Stone attempted to persuade the International Music Institute Darmstadt to cooperate and, following on from the debates there, to systematically record various forms of notation together. In a second step, the content of the debates at the conference is examined, with a particular focus on the different and sometimes conflicting perspectives of interpreters and composers. Numerous connections to fundamental aesthetic discussions of the time can be worked out, in particular to the relationship between the composer’s intention and interpretation, which was renegotiated in a form of notation that was individualized to the extreme. Finally, with a view to later discussions, this topic is pointed to the question of the relationship between morphology and musical structure, exemplified by positions of Wolfgang Rihm (1982), Klaus Huber (1988) and John Cage (1990).

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