scholarly journals Expression and Secretion of an Atrial Natriuretic Peptide in Beige-Like 3T3-L1 Adipocytes

2019 ◽  
Vol 20 (24) ◽  
pp. 6128 ◽  
Author(s):  
In-Seon Bae ◽  
Sang Hoon Kim
Keyword(s):  
Atrial Natriuretic Peptide ◽  
Natriuretic Peptide ◽  
Peptide Hormone ◽  
Brown Adipocyte ◽  
Omega 3 ◽  
Hek 293 ◽  
Hek 293 Cells ◽  
293 Cells ◽  
Beige Adipocytes ◽  
Atrial Natriuretic

The browning of white adipose tissue (beige adipocytes) stimulates energy expenditure. Omega-3 fatty acids have been shown to induce thermogenic action in adipocytes via G-protein coupled receptor 120 (GPR120). Atrial natriuretic peptide (ANP) is a peptide hormone that plays the role of maintaining normal blood pressure in kidneys to inhibit Na+ reuptake. Recently, ANP was found to induce adipocyte browning by binding to NPR1, an ANP receptor. However, the expression of ANP in adipocytes has not yet been studied. Therefore, in this study, we investigate the expression of ANP in beige-like adipocytes induced by docosahexaenoic acids (DHA), T3, or a PPAR agonist, rosiglitazone. First, we found that brown adipocyte-specific genes were upregulated in beige-like adipocytes. DHA promoted ANP expression in beige-like cells, whereas DHA-induced ANP expression was abolished by GPR120 knockout. ANP secretion of beige-like adipocytes was increased via PKC/ERK1/2 signaling in the GPR120 pathway. Furthermore, ANP secreted from beige-like adipocytes acted on HEK-293 cells, the recipient cells, leading to increased cGMP activity. After the NPR1 knockdown of HEK-293 cells, cGMP activity was not changed. Taken together, our findings indicate that beige-like adipocytes induce ANP secretion, which may contribute to improving obesity-associated metabolic disease.

scholarly journals Role of Glycosylation in Corin Zymogen Activation

2007 ◽  
Vol 282 (38) ◽  
pp. 27728-27735 ◽  
Author(s):  
Xudong Liao ◽  
Wei Wang ◽  
Shenghan Chen ◽  
Qingyu Wu
Keyword(s):  
Atrial Natriuretic Peptide ◽  
Natriuretic Peptide ◽  
Cysteine Proteases ◽  
Biologically Active ◽  
Zymogen Activation ◽  
Hek 293 ◽  
Hek 293 Cells ◽  
Neonatal Cardiomyocytes ◽  
293 Cells ◽  
Atrial Natriuretic

The cardiac serine protease corin is the pro-atrial natriuretic peptide convertase. Corin is made as a zymogen, which is activated by proteolytic cleavage. Previous studies showed that recombinant human corin expressed in HEK 293 cells was biologically active, but activated corin fragments were not detectable, making it difficult to study corin activation. In this study, we showed that recombinant rat corin was activated in HEK 293 cells, murine HL-1 cardiomyocytes, and rat neonatal cardiomyocytes. In these cells, activated corin represented a small fraction of the total corin molecules. The activation of recombinant rat corin was inhibited by small molecule trypsin inhibitors but not inhibitors for matrix metalloproteinases or cysteine proteases, suggesting that a trypsin-like protease activated corin in these cells. Glycosidase digestion showed that rat and human corin proteins contained substantial N-glycans but little O-glycans. Treatment of HEK 293 cells expressing rat corin with tunicamycin prevented corin activation and inhibited its pro-atrial natriuretic peptide processing activity. Similar effects of tunicamycin on endogenous corin activity were found in HL-1 cells. Mutations altering the two N-glycosylation sites in the protease domain of rat corin prevented its activation in HEK 293 and HL-1 cells. Our results indicate that N-linked oligosaccharides play an important role in corin activation.

Pro-atrial natriuretic peptide hormone from right atria is correlated with cardiac depression in septic patients

2001 ◽  
Vol 24 (7) ◽  
pp. RC22-RC24 ◽  
Author(s):  
Branka Mazul-Sunko ◽  
N. Zarkovic ◽  
N. Vrkic ◽  
R. Klinger ◽  
M. Peric ◽  
...  
Keyword(s):  
Atrial Natriuretic Peptide ◽  
Natriuretic Peptide ◽  
Peptide Hormone ◽  
Cardiac Depression ◽  
Right Atria ◽  
Atrial Natriuretic

scholarly journals Processing of Pro–B-Type Natriuretic Peptide: Furin and Corin as Candidate Convertases2

2010 ◽  
Vol 56 (7) ◽  
pp. 1166-1176 ◽  
Author(s):  
Alexander G Semenov ◽  
Natalia N Tamm ◽  
Karina R Seferian ◽  
Alexander B Postnikov ◽  
Natalia S Karpova ◽  
...  
Keyword(s):  
Heart Failure ◽  
Natriuretic Peptide ◽  
Spectrometric Analysis ◽  
Precursor Molecule ◽  
Hek 293 ◽  
Hek 293 Cells ◽  
293 Cells ◽  
Lovo Cells ◽  
Interfering Rna

Abstract Background: B-type natriuretic peptide (BNP) and its N-terminal fragment (NT-proBNP) are the products of the enzyme-mediated cleavage of their precursor molecule, proBNP. The clinical significance of proBNP-derived peptides as biomarkers of heart failure has been explored thoroughly, whereas little is known about the mechanisms of proBNP processing. We investigated the role of 2 candidate convertases, furin and corin, in human proBNP processing. Methods: We measured proBNP expression in HEK 293 and furin-deficient LoVo cells. We used a furin inhibitor and a furin-specific small interfering RNA (siRNA) to explore the implication of furin in proBNP processing. Recombinant proBNPs were incubated with HEK 293 cells transfected with the corin-expressing plasmid. We applied mass spectrometry to analyze the products of furin- and corin-mediated cleavage. Results: Reduction of furin activity significantly impaired proBNP processing in HEK 293 cells. Furin-deficient LoVo cells were unable to process proBNP, whereas coexpression with furin resulted in effective proBNP processing. Mass spectrometric analysis revealed that the furin-mediated cleavage of proBNP resulted in BNP 1–32, whereas corin-mediated cleavage led to the production of BNP 4–32. Some portion of proBNP in the plasma of heart failure patients was not glycosylated in the cleavage site region and was susceptible to furin-mediated cleavage. Conclusions: Both furin and corin are involved in the proBNP processing pathway, giving rise to distinct BNP forms. The significance of the presence of unprocessed proBNP in circulation that could be cleaved by the endogenous convertases should be further investigated for better understanding BNP physiology.

HCaRG is a novel regulator of renal epithelial cell growth and differentiation causing G2M arrest

2003 ◽  
Vol 284 (4) ◽  
pp. F753-F762 ◽  
Author(s):  
Alison M. Devlin ◽  
Nicolas Solban ◽  
Sandra Tremblay ◽  
Jolanta Gutkowska ◽  
Walter Schürch ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Growth ◽  
Epithelial Cell ◽  
Atrial Natriuretic Peptide ◽  
Cell Size ◽  
Natriuretic Peptide ◽  
Renal Epithelial Cell ◽  
Hek 293 ◽  
Cell Growth And Differentiation ◽  
Atrial Natriuretic

We recently identified a novel calcium-regulated gene, HCaRG, that is highly expressed in the kidney and maps to a chromosomal locus determining kidney weight in rats. The mRNA levels of HCaRG negatively correlate with the proliferative status of the kidney cells. To investigate its role in renal epithelial cellular growth directly, we studied the human embryonic kidney cell line (HEK-293) stably transfected with either plasmid alone or plasmid containing rat HCaRG. [3H]thymidine incorporation was significantly lower in HCaRG clones. Although HCaRG clones exhibited some enhanced susceptibility to cell death, this was not the primary mechanism of reduced proliferation. Cell cycle analysis revealed a G2M phase accumulation in HCaRG clones that was associated with upregulation of p21Cip1/WAF1 and downregulation of p27Kip1. HCaRG clones had a greater protein content, larger cell size, and released 4.5- to 8-fold more of an atrial natriuretic peptide-like immunoreactivity compared with controls. In addition, HCaRG clones demonstrated the presence of differentiated junctions and a lower incidence of mitotic figures. Genistein treatment of wild-type HEK-293 cells mimicked several phenotypic characteristics associated with HCaRGoverexpresssion, including increased cell size and increased release of atrial natriuretic peptide. Taken together, our results suggest that HCaRG is a regulator of renal epithelial cell growth and differentiation causing G2M cell cycle arrest.

scholarly journals Interaction between bradykinin and natriuretic peptides via RGS protein activation in HEK-293 cells

2012 ◽  
Vol 303 (12) ◽  
pp. C1260-C1268 ◽  
Author(s):  
Marina Dobrivojević ◽  
Aleksandra Sinđić ◽  
Bayram Edemir ◽  
Stefanie Kalweit ◽  
Wolf-Georg Forssmann ◽  
...  
Keyword(s):  
Signaling Pathway ◽  
Natriuretic Peptide ◽  
Natriuretic Peptides ◽  
Specific Inhibitor ◽  
Rgs Proteins ◽  
Patch Clamp Technique ◽  
Hek 293 ◽  
Hek 293 Cells ◽  
293 Cells ◽  
Intracellular Ca

In this study, the interaction of natriuretic peptides (NP) and bradykinin (BK) signaling pathways was identified by measuring membrane potential ( Vm) and intracellular Ca2+ using the patch-clamp technique and flow cytometry in HEK-293 cells. BK and NP receptor mRNA was identified using RT-PCR. BK (100 nM) depolarized cells activating bradykinin receptor type 2 (B2R) and Ca2+-dependent Cl− channels inhibitable by 5-nitro-2-(3-phenylpropylamino)benzoic acid (NPPB; 10 μM). The BK-induced Ca2+ signal was blocked by the B2R inhibitor HOE 140. [Des-Arg9]-bradykinin, an activator of B1R, had no effect on intracellular Ca2+. NP [atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), C-type natriuretic peptide (CNP), and urodilatin] depolarized HEK-293 cells inhibiting K+ channels. ANP, urodilatin, BNP [binding to natriuretic peptide receptor (NPR)-A] and 8-bromo-(8-Br)-cGMP inhibited the BK-induced depolarization while CNP (binding to NPR-Bi) failed to do so. The inhibitory effect on BK-triggered depolarization could be reversed by blocking PKG using the specific inhibitor KT 5823. BK-stimulated depolarization as well as Ca2+ signaling was completely blocked by the phospholipase C (PLC) inhibitor U-73122 (10 nM). The inositol 1,4,5-trisphosphate receptor blocker 2-aminoethoxydiphenyl borate (2-APB; 50 μM) completely inhibited the BK-induced Ca2+ signaling. UTP, another activator of the PLC-mediated Ca2+ signaling pathway, was blocked by U-73122 as well but not by 8-Br-cGMP, indicating an intermediate regulatory step for NP via PKG in BK signaling such as regulators of G-protein signaling (RGS) proteins. When RGS proteins were inhibited by CCG-63802 in the presence of BK and 8-Br-cGMP, cells started to depolarize again. In conclusion, as natural antagonists of the B2R signaling pathway, NP may also positively interact in pathological conditions caused by BK.

scholarly journals An Activating Deletion Variant in the Submembrane Region of Natriuretic Peptide Receptor-B Causes Tall Stature

2020 ◽  
Vol 105 (7) ◽  
pp. 2354-2366 ◽  
Author(s):  
Peter Lauffer ◽  
Erick Miranda-Laferte ◽  
Hermine A van Duyvenvoorde ◽  
Arie van Haeringen ◽  
Franziska Werner ◽  
...  
Keyword(s):  
Natriuretic Peptide ◽  
Skin Fibroblasts ◽  
Growth Disorders ◽  
Tall Stature ◽  
Natriuretic Peptide Receptor ◽  
Hek 293 ◽  
Peptide Receptor ◽  
Cgmp Production ◽  
Hek 293 Cells ◽  
293 Cells

Abstract Context C-type natriuretic peptide (CNP) is critically involved in endochondral bone growth. Variants in the genes encoding CNP or its cyclic guanosine monophosphate (cGMP)-forming receptor (natriuretic peptide receptor-B [NPR-B], gene NPR2) cause monogenic growth disorders. Here we describe a novel gain-of-function variant of NPR-B associated with tall stature and macrodactyly of the great toes (epiphyseal chondrodysplasia, Miura type). Design History and clinical characteristics of 3 family members were collected. NPR2 was selected for sequencing. Skin fibroblasts and transfected HEK-293 cells were used to compare mutant versus wild-type NPR-B activities. Homology modeling was applied to understand the molecular consequences of the variant. Results Mother’s height was +2.77 standard deviation scores (SDS). The heights of her 2 daughters were +1.96 SDS at 7 years and +1.30 SDS at 4 years of age. Skeletal surveys showed macrodactyly of the great toes and pseudo-epiphyses of the mid- and proximal phalanges. Sequencing identified a novel heterozygous variant c.1444_1449delATGCTG in exon 8 of NPR2, predicted to result in deletion of 2 amino acids Met482-Leu483 within the submembrane region of NPR-B. In proband’s skin fibroblasts, basal cGMP levels and CNP-stimulated cGMP production were markedly increased compared with controls. Consistently, assays with transfected HEK-293 cells showed markedly augmented baseline and ligand-dependent activity of mutant NPR-B. Conclusions We report the second activating variant within the intracellular submembrane region of NPR-B resulting in tall stature and macrodactyly. Our functional and modeling studies suggest that this domain plays a critical role in the baseline conformation and ligand-dependent structural rearrangement of NPR-B required for cGMP production.

Corin, a Transmembrane Cardiac Serine Protease, as a Pro-Atrial Natriuretic Peptide Convertase

Hypertension ◽  
2000 ◽  
Vol 36 (suppl_1) ◽  
pp. 709-710
Author(s):  
Wei Yan ◽  
Faye Wu ◽  
John Morser ◽  
Qingyu Wu
Keyword(s):  
Blood Pressure ◽  
Atrial Natriuretic Peptide ◽  
Serine Protease ◽  
Natriuretic Peptide ◽  
Transmembrane Protein ◽  
Biologically Active ◽  
Expression Vectors ◽  
Sequence Specificity ◽  
293 Cells ◽  
Atrial Natriuretic

P92 Atrial natriuretic peptide (ANP) is a cardiac hormone that reduces high blood pressure by promoting salt excretion, decreasing blood volume and relaxing vessel tension. It is implicated in major cardiovascular diseases such as hypertension and congestive heart failure. In cardiomyocytes, ANP is synthesized as a precursor, pro-ANP, that is converted to biologically active ANP by an unknown membrane-associated serine protease. Recently, we cloned a novel cardiac serine protease, corin, that has the predicted structure of a type II transmembrane protein. Northern and in situ hybridization detected corin mRNA in the heart where its expression was most abundant in cardiomyocytes of the atrium. Corin mRNA was also detected in developing kidney and bones. The overall expression pattern of corin mRNA was very similar to that of ANP, leading to our hypothesis that corin is the pro-ANP convertase. To test this hypothesis, we constructed expression vectors for both human corin and pro-ANP. Recombinant corin and pro-ANP were expressed in human 293 cells. Effects of corin on pro-ANP processing was examined by western analysis. The results showed that human corin, but not control proteases prothrombin and hepsin, converted pro-ANP to ANP in the cell-based experiments. To determine sequence specificity of corin-mediated cleavage in pro-ANP, we constructed a mutant pro-ANP R98G in which the predicted cleave site residue Arg98 was replaced with a Gly. In co-transfection experiments, corin processed wild-type pro-ANP but not mutant pro-ANP R98G, indicating that corin-mediated cleavage in pro-ANP is highly sequence specific. We also examined effects of protease inhibitors on processing of pro-ANP by corin. Our results showed that the activity of corin was inhibited by aprotinin, benzamidine and leupeptin but not soybean trypsin inhibitor. Similar effects of these proteases on pro-ANP processing were reported previously. Thus, corin matches all known characteristics of the long-sought pro-ANP convertase. Identification of corin as the pro-ANP convertase suggests a new regulatory mechanism for the ANP-mediated pathway that is important for controlling blood pressure.

scholarly journals Regulation of atrial natriuretic peptide (ANP) and its role in blood pressure

2012 ◽  
Vol 1 (7) ◽  
pp. 176-179 ◽  
Author(s):  
Feroz Ahmed ◽  
Nahida Tabassum ◽  
Saima Rasool
Keyword(s):  
Heart Failure ◽  
Blood Pressure ◽  
Atrial Natriuretic Peptide ◽  
Natriuretic Peptide ◽  
Renin Angiotensin System ◽  
Peptide Hormone ◽  
Decompensated Heart Failure ◽  
Pharmaceutical Journal ◽  
The Body ◽  
Atrial Natriuretic

Atrial natriuretic peptide (ANP) a powerful vasodilator, and a protein (28-amino acid peptide) hormone secreted by heart muscle cells. It is released in response to atrial distention, stretching of the vessel walls, sympathetic stimulation of ?-adrenoceptors, raised sodium concentration, angiotensin-II and endothelin. ANP binds to three cell surface receptors called ANP receptors. The overall effect of ANP on the body is to counter increases in blood pressure and volume caused by the renin-angiotensin system. It has also been reported to increase the release of free fatty acids from adipose tissue. Regulation of its effects is achieved through gradual degradation of the peptide by the enzyme neutral endopeptidase (NEP). Inhibitors of NEP are currently being developed to treat disorders ranging from hypertension to heart failure. Synthetic analogs of ANP have been investigated as potential therapies for the treatment of decompensated heart failure and other diseases.DOI: http://dx.doi.org/10.3329/icpj.v1i7.10812International Current Pharmaceutical Journal 2012, 1(7): 176-179 

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