Maturation of cholecystokinin receptors in pancreatic acini of rats

1986 ◽  
Vol 250 (5) ◽  
pp. G594-G597 ◽  
Author(s):  
Y. K. Leung ◽  
P. C. Lee ◽  
E. Lebenthal
Keyword(s):  
Receptor Binding ◽  
Binding Capacity ◽  
Amylase Secretion ◽  
Newborn Rat ◽  
Pancreatic Acini ◽  
Binding System ◽  
High Affinity ◽  
Cholecystokinin Receptors ◽  
Maximal Binding

Amylase secretion by fetal and newborn rat pancreata was shown not to respond to cholecystokinin (CCK) stimulation. Binding of 125I-Bolton-Hunter (125I-BH)-CCK-8 to dispersed pancreatic acini from rats at various perinatal ages was measured and compared with adults. Binding occurred maximally at 30 min at 37 degrees C at all ages. Maximal binding increased with age. The capacity and affinity of the CCK-receptor binding system was analyzed by Scatchard's method. The results revealed that the capacity of the high-affinity component gradually increases with age, whereas that of the low-affinity component remains relatively constant. Amylase secretion due to CCK stimulation also increases with age, suggesting that the lack of responsiveness of pancreata of pups to secretagogue is due to a low binding capacity of the high-affinity component of the receptors.

Regulation of bombesin receptors on pancreatic acini by cholecystokinin

1989 ◽  
Vol 256 (2) ◽  
pp. G291-G298
Author(s):  
M. Younes ◽  
S. A. Wank ◽  
R. Vinayek ◽  
R. T. Jensen ◽  
J. D. Gardner
Keyword(s):  
Guinea Pig ◽  
Response Curve ◽  
Binding Capacity ◽  
The Other ◽  
Single Class ◽  
Pancreatic Acini ◽  
Cck Receptors ◽  
Cyclic Monophosphate ◽  
High Affinity ◽  
Bombesin Receptors

When guinea pig pancreatic acini are first incubated with the COOH-terminal octapeptide of cholecystokinin (CCK-8), washed, and then reincubated with 125I-[Tyr4]bombesin (125I-[Tyr4]BN) there is a significant decrease in binding of 125I-[Tyr4]BN compared with that observed with pancreatic acini that have been first incubated with no additions. The CCK-8-induced decrease in binding is maximal after 90 min of first incubation is abolished by reducing the temperature of the first incubation from 37 to 4 degrees C or by adding L364,718 to the first incubation and cannot be reproduced by first incubating acini with A23187, 8-bromoadenosine 3',5'-cyclic monophosphate (8Br-cAMP), 8-bromoadenosine 3',5'-cyclic monophosphate (8Br-cGMP), or 12-O-tetradecanoylphorbol 13-acetate. 125I-[Tyr4]BN interacts with a single class of receptors on pancreatic acini, and first incubating acini with CCK-8 decreases the affinity of BN receptors for BN with no change in the maximal binding capacity. CCK-8 does not alter the rate at which bound 125I-[Tyr4]BN dissociates from pancreatic acini; therefore, CCK-8 must alter the rate at which the radiolabeled BN analogue associates with its receptor. Pancreatic acini possess two classes of CCK receptors: one has a high affinity for CCK-8; the other has a low affinity for CCK-8. The dose-response curve for CCK-8-induced inhibition of binding of 125I-[Tyr4]BN appears to to reflect occupation of low-affinity CCK receptors by CCK-8.(ABSTRACT TRUNCATED AT 250 WORDS)

Cholecystokinin-induced persistent stimulation of enzyme secretion from pancreatic acini

1981 ◽  
Vol 240 (6) ◽  
pp. G459-G465 ◽  
Author(s):  
S. M. Collins ◽  
S. Abdelmoumene ◽  
R. T. Jensen ◽  
J. D. Gardner
Keyword(s):  
Enzyme Secretion ◽  
Amylase Secretion ◽  
Intermediate Step ◽  
Pancreatic Acini ◽  
Persistent Effect ◽  
Incubation Solution ◽  
Cholecystokinin Receptors ◽  
Incomplete Removal ◽  
High Concentrations ◽  
Stimulation Of

When pancreatic acini are first incubated with cholecystokinin, washed to remove free cholecystokinin and then reincubated in fresh incubation solution, there is significant residual stimulation of amylase secretion. This residual stimulation requires relatively high concentrations of the secretagogue, is reversible, and is specific for cholecystokinin. Induction of residual stimulation occurs more rapidly at 37 degrees C (maximal by 1 min) than at 4 degrees C (maximal by 10 min), and, once induced, residual stimulation persists for up to 75 min at 37 degrees C and for more than 90 min at 4 degrees C. The persistent effect of cholecystokinin on enzyme secretion cannot be accounted for by incomplete removal of the secretagogue by the wash procedure or by activation of some intermediate step in the mechanism of action of cholecystokinin that persists after the secretagogue dissociates from its receptors. Instead, cholecystokinin-induced residual stimulation of enzyme secretion appears to result from persistent occupation of cholecystokinin receptors by the secretagogue.

Somatostatin binding to chicken adenohypophysial membranes

1990 ◽  
Vol 4 (3) ◽  
pp. 213-221 ◽  
Author(s):  
S. Harvey ◽  
D. Attardo ◽  
J. S. Baidwan
Keyword(s):  
Binding Sites ◽  
Binding Capacity ◽  
Caudal Lobe ◽  
Gh Secretion ◽  
High Affinity ◽  
Cephalic Lobe ◽  
Pituitary Glands ◽  
Lh Release ◽  
Maximal Binding ◽  
Maximal Suppression

ABSTRACT [125I-Tyr1]-Somatostatin (SRIF)-binding sites were demonstrated on crude plasma membrane preparations from chicken pituitary glands. These binding sites were saturable and of high affinity (dissociation constant <1·0 nm) and low capacity (maximal binding capacity <200 fmol/mg protein) and were specific for SRIF moieties. The number and affinity of these binding sites in the caudal lobe of the pituitary, in which somatotrophs predominate, were similar to those in the cephalic lobe, in which lactotrophs and thyrotrophs are confined. Gonadotrophs are present in the caudal lobe, but whereas exogenous SRIF inhibited secretagogue-induced GH release from incubated pituitary glands, it had no effect on basal or secretagogue-induced LH release. The half-maximal binding of SRIF to the caudal lobe membranes (3 nm) was similar to that required for half-maximal suppression of TRH-induced GH release, suggesting a role for these binding sites in the regulation of GH secretion in birds.

Binding of somatostatin to pancreatic acinar cells

1984 ◽  
Vol 247 (1) ◽  
pp. G62-G69 ◽  
Author(s):  
J. P. Esteve ◽  
C. Susini ◽  
N. Vaysse ◽  
H. Antoniotti ◽  
E. Wunsch ◽  
...  
Keyword(s):  
Binding Sites ◽  
Binding Capacity ◽  
Somatostatin Receptors ◽  
Subcellular Fractionation ◽  
Membrane Receptors ◽  
Pancreatic Acinar Cells ◽  
Scatchard Analysis ◽  
Single Class ◽  
Pancreatic Acini ◽  
Maximal Binding

The binding of 125I-[Tyr11]somatostatin to guinea pig pancreatic acini was saturable and temperature, protein, and radioligand concentration dependent. Dissociation rate was very slow (t1/2 = 193 +/- 24 min). Scatchard analysis revealed a single class of binding sites with a Kd of 0.28 +/- 0.02 nM and a maximal binding capacity of 72 +/- 10.6 fmol/mg prot. There was a strong correlation between binding capacity and extracellular calcium concentration. Incubating acini in EGTA-containing medium with no added Ca2+ caused a 50% decrease in maximal binding capacity with a decrease in receptor affinity. Furthermore, in the absence of calcium, bound somatostatin was rapidly released (t1/2 = 14 +/- 1 min). Subcellular fractionation studies and acid treatment of acini incubated with the tracer showed that most of the somatostatin binding sites were located on the cell surface. Agents that altered cellular calcium in pancreatic acini, such as analogues of cholecystokinin and cholinergic agents, also inhibited the binding of 125I-[Tyr11]somatostatin by a calcium-dependent process. We conclude that somatostatin binds to specific plasma membrane receptors to form a slowly reversible complex that is highly reactive with calcium. Cell calcium-mobilizing agents decrease the affinity of acinar somatostatin receptors for somatostatin.

High-affinity CCK receptors are coupled to phospholipase A2 pathways to mediate pancreatic amylase secretion

1995 ◽  
Vol 269 (3) ◽  
pp. G435-G444 ◽  
Author(s):  
Y. Tsunoda ◽  
C. Owyang
Keyword(s):  
Phospholipase A2 ◽  
Acinar Cells ◽  
Receptor Activation ◽  
Pancreatic Acinar Cells ◽  
Rabbit Serum ◽  
Amylase Secretion ◽  
Pancreatic Acini ◽  
Cck Receptors ◽  
Permeabilized Cells ◽  
High Affinity

It is well recognized that JMV-180, a cholecystokinin (CCK) analogue, acts as an agonist on the high-affinity CCK receptor in pancreatic acinar cells. It caused Ca2+ oscillations and amylase secretion in a manner independent of the phospholipase C-inositol 1,4,5-trisphosphate (IP3) pathway. We investigated the mechanism by which the high-affinity CCK receptor utilizes IP3-independent Ca2+ signal transduction to mediate amylase secretion. JMV-180 (1-1,000 nM)-stimulated Ca2+ oscillations and amylase secretion were significantly inhibited by the phospholipase A2 (PLA2) inhibitor, ONO-RS-082 (10 microM). Using streptolysin O-permeabilized cells, we showed that a porcine pancreatic anti-PLA2 antibody from rabbit serum (250 ng/ml) inhibited JMV-180-stimulated amylase secretion. In contrast to CCK octapeptide, JMV-180 (1 nM-10 microM) had no effect on intracellular IP3 levels. These concentrations of JMV-180 did, however, increase intracellular levels of arachidonic acid (AA) metabolite by 2.5-fold in a biphasic manner. Application of exogenous AA (10 microM) released 60% of ATP-incorporated 45Ca2+ from permeabilized pancreatic acini within 3 min in a transient manner. We also showed that active phorbol ester (100 nM) inhibited Ca2+ oscillations and amylase secretion stimulated by JMV-180 (10 nM) or CCK-OPE (100 nM). Application of Mn2+ (2 mM) to superfused acini resulted in a rapid quench of fura 2 fluorescence during 10 nM JMV-180 stimulation, suggesting an involvement of extracellular Ca2+ influx. However, the major source of Ca2+ utilized for oscillations during high-affinity CCK receptor activation was intracellular. In conclusion, we have demonstrated that the high-affinity CCK receptors are coupled to PLA2 pathways to produce AA, which mediates cytosolic Ca2+ oscillation and monophasic amylase secretion, in rat pancreatic acinar cells.

Carbachol acts through protein kinase C to modulate cholecystokinin receptors on pancreatic acini

1991 ◽  
Vol 261 (6) ◽  
pp. G981-G986 ◽  
Author(s):  
D. S. Louie ◽  
O. Y. Chung
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Binding Capacity ◽  
Muscarinic Agonist ◽  
Amylase Release ◽  
Pancreatic Acini ◽  
Cck Receptors ◽  
High Affinity ◽  
Total Binding ◽  
Kinase C

Cholecystokinin (CCK) and cholinergic agonists are both major stimulants of pancreatic enzyme secretion and both utilize a common calcium-phosphoinositide-mediated receptor coupling system. In this study we investigated the modulation of pancreatic acinar CCK receptors by the muscarinic agonist carbachol (CCh) and investigated the intracellular mechanisms involved in the modulation. Acini were isolated from rat pancreas and dispersed in N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid-Ringer solution. Preincubation with 0.1 mM carbachol for 60 min reduced the CCK octapeptide (CCK-8; 100 pM)-stimulated amylase release by 43 +/- 5%. Binding of 125I-Bolton-Hunter-labeled CCK-8 (125I-BH-CCK-8) revealed two classes of CCK receptors, a high affinity with a dissociation constant (Kd) of 20 pM and a low affinity with a Kd of 2.3 nM. Pretreatment with 100 microM CCh decreased total binding by 35 +/- 6%, affecting the binding capacity of the high-affinity site, without change in the maximal binding capacity of the low-affinity site and no change in the Kd of either site. Preincubation of acini with 12-O-tetradecanoylphorbol 12,13-acetate (TPA, 1 microM), an activator of protein kinase C (PKC), decreased subsequent CCK-8-stimulated amylase release, and total binding of 125I-BH-CCK-8 to a similar extent as with pretreatment with CCh. The inhibitory effect of TPA or CCh on CCK-8-stimulated amylase release was reversed by simultaneous preincubation with H-7, an inhibitor of PKC. Pretreatment of acini with the calcium ionophore A23187, vasoactive intestinal peptide, or 8-bromoadenosine 3',5'-cyclic monophosphate had no effect on 125I-BH-CCK-8 binding. After CCh or TPA preincubation, CCK-8-stimulated production of [3H]inositol phosphates was inhibited by at least 49%.(ABSTRACT TRUNCATED AT 250 WORDS)

Receptors for vasoactive intestinal peptide and secretin on rat pancreatic acini

1984 ◽  
Vol 246 (6) ◽  
pp. G710-G717 ◽  
Author(s):  
B. M. Bissonnette ◽  
M. J. Collen ◽  
H. Adachi ◽  
R. T. Jensen ◽  
J. D. Gardner
Keyword(s):  
Vasoactive Intestinal Peptide ◽  
Binding Sites ◽  
Enzyme Secretion ◽  
Amylase Secretion ◽  
Temperature Dependent ◽  
Pancreatic Acini ◽  
Rat Pancreas ◽  
High Affinity ◽  
Fourth Class ◽  
Stimulation Of

In dispersed acini from rat pancreas, binding of 125I-labeled vasoactive intestinal peptide and 125I-labeled secretin was relatively rapid, reversible, saturable, and temperature dependent. The rate of dissociation of bound 125I-labeled peptide was not a function of the concentration of free vasoactive intestinal peptide or secretin, indicating that the apparent affinities of these labeled peptides for their binding sites do not depend on the extent of receptor occupation. Four classes of receptors are required to account for the actions of vasoactive intestinal peptide and secretin on enzyme secretion, cellular cAMP, and binding of 125I-vasoactive intestinal peptide and 125I-secretin. One class has a high affinity for vasoactive intestinal peptide, and occupation of this class of receptors causes increased cellular cAMP and stimulation of amylase secretion. A second class has a low affinity for vasoactive intestinal peptide and for secretin, and occupation of these receptors does not cause changes in cAMP or amylase secretion. A third class of receptors has a high affinity for secretin, and occupation of these receptors causes increased cAMP and stimulation of amylase secretion. A fourth class of receptors has a low affinity for secretin, and occupation of these receptors causes stimulation of amylase secretion by a non-cAMP-mediated mechanism.

Characterization of gastrin receptors on guinea pig pancreatic acini

1987 ◽  
Vol 253 (6) ◽  
pp. G793-G801 ◽  
Author(s):  
D. H. Yu ◽  
M. Noguchi ◽  
Z. C. Zhou ◽  
M. L. Villanueva ◽  
J. D. Gardner ◽  
...  
Keyword(s):  
Guinea Pig ◽  
Binding Capacity ◽  
Pancreatic Tumors ◽  
Amylase Secretion ◽  
Single Class ◽  
Amylase Release ◽  
Pancreatic Acini ◽  
Cyclic Monophosphate ◽  
Relative Potencies

Recent studies have demonstrated gastrin receptors in some pancreatic tumors and that gastrin is a potent stimulant of pancreatic Na+-H+ exchange. In the present study we used 125I-labeled gastrin (125I-gastrin) to characterize gastrin receptors on guinea pig pancreatic acini. Binding of 125I-gastrin was temperature dependent, saturable, and specific for gastrin-related peptides. Analysis demonstrated a single class of receptors with high affinity for gastrin (Kd = 1.5 nM) and a binding capacity of 1 fmol/mg protein. Binding of 125I-gastrin was inhibited with the following relative potencies (Kd): cholecystokinin octapeptide (CCK-8) (0.35 nM) greater than gastrin-17-I = gastrin-34-I (1.5 nM) greater than pentagastrin (7 nM) greater than desulfated [des(SO3)]CCK-8 (28 nM) greater than CCK-4 (508 nM) and by the receptor antagonists CBZ-CCK-27-32-NH2 (3.5 microM) greater than proglumide analogue 10 (30 microM) greater than asperlicin (265 microM) greater than Bt2-guanosine 3',5'-cyclic monophosphate (828 micron). In contrast, for both stimulation of enzyme secretion and inhibition of binding of 125I-CCK-8 the relative potencies were CCK-8 much greater than des(SO3)CCK-8 greater than gastrin-17-I = gastrin-34-I greater than pentagastrin greater than CCK-4. For each receptor antagonist the dose-inhibition curve for gastrin-stimulated amylase release was superimpossible with that for CCK-8-stimulated amylase release. Gastrin-17-I at concentrations less than 0.1 microM did not potentiate carbachol or vasoactive intestinal peptide-stimulated amylase secretion and did not affect basal or stimulated adenosine 3',5'-cyclic monophosphate or 45Ca outflux.(ABSTRACT TRUNCATED AT 250 WORDS)

Is the Rigidity of SARS-CoV-2 Spike Receptor-Binding Motif the Hallmark for Its Enhanced Infectivity and Pathogenesis?

2020 ◽  
Author(s):  
Angelo Spinello ◽  
Andrea Saltalamacchia ◽  
Alessandra Magistrato
Keyword(s):  
Health Economic ◽  
Global Health ◽  
Receptor Binding ◽  
Binding Motif ◽  
Spike Glycoprotein ◽  
High Affinity ◽  
Global Pandemic ◽  
Medical Countermeasures ◽  
High Infectivity ◽  
Human Receptor

<p>The latest outbreak of a new pathogenic coronavirus (SARS-CoV-2) is provoking a global health, economic and societal crisis. All-atom simulations enabled us to uncover the key molecular traits underlying the high affinity of SARS-CoV-2 spike glycoprotein towards its human receptor, providing a rationale to its high infectivity. Harnessing this knowledge can boost developing effective medical countermeasures to fight the current global pandemic.</p>

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