Arg13 of B-Type Natriuretic Peptide Reciprocally Modulates Binding to Guanylyl Cyclase but Not Clearance Receptors

Dickey, Deborah M.; Barbieri, Kathryn A; McGuirk, C. Michael; Potter, Lincoln R.

doi:10.1124/mol.110.066084

Cited by 10 publications

(6 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…These intracellular cGMP measurements have limited accuracy, precision and reproducibility. They are tedious with the preparation of cell lysate and depend on standard curve by radioimmunoassays [ 206 ]. One stable cell-based assay is based on the NPR-A in HEK 293 cell.…”

Section: Diagnostic Valuesmentioning

confidence: 99%

Synthesis, secretion, function, metabolism and application of natriuretic peptides in heart failure

et al. 2018

View full text Add to dashboard Cite

As a family of hormones with pleiotropic effects, natriuretic peptide (NP) system includes atrial NP (ANP), B-type NP (BNP), C-type NP (CNP), dendroaspis NP and urodilatin, with NP receptor-A (guanylate cyclase-A), NP receptor-B (guanylate cyclase-B) and NP receptor-C (clearance receptor). These peptides are genetically distinct, but structurally and functionally related for regulating circulatory homeostasis in vertebrates. In humans, ANP and BNP are encoded by NP precursor A (NPPA) and NPPB genes on chromosome 1, whereas CNP is encoded by NPPC on chromosome 2. NPs are synthesized and secreted through certain mechanisms by cardiomyocytes, fibroblasts, endotheliocytes, immune cells (neutrophils, T-cells and macrophages) and immature cells (embryonic stem cells, muscle satellite cells and cardiac precursor cells). They are mainly produced by cardiovascular, brain and renal tissues in response to wall stretch and other causes. NPs provide natriuresis, diuresis, vasodilation, antiproliferation, antihypertrophy, antifibrosis and other cardiometabolic protection. NPs represent body's own antihypertensive system, and provide compensatory protection to counterbalance vasoconstrictor-mitogenic-sodium retaining hormones, released by renin-angiotensin-aldosterone system (RAAS) and sympathetic nervous system (SNS). NPs play central roles in regulation of heart failure (HF), and are inactivated through not only NP receptor-C, but also neutral endopeptidase (NEP), dipeptidyl peptidase-4 and insulin degrading enzyme. Both BNP and N-terminal proBNP are useful biomarkers to not only make the diagnosis and assess the severity of HF, but also guide the therapy and predict the prognosis in patients with HF. Current NP-augmenting strategies include the synthesis of NPs or agonists to increase NP bioactivity and inhibition of NEP to reduce NP breakdown. Nesiritide has been established as an available therapy, and angiotensin receptor blocker NEP inhibitor (ARNI, LCZ696) has obtained extremely encouraging results with decreased morbidity and mortality. Novel pharmacological approaches based on NPs may promote a therapeutic shift from suppressing the RAAS and SNS to re-balancing neuroendocrine dysregulation in patients with HF. The current review discussed the synthesis, secretion, function and metabolism of NPs, and their diagnostic, therapeutic and prognostic values in HF.

show abstract

Section: Diagnostic Valuesmentioning

confidence: 99%

Synthesis, secretion, function, metabolism and application of natriuretic peptides in heart failure

et al. 2018

View full text Add to dashboard Cite

show abstract

“…Phosphorylation is required for peptide activation, and prolonged natriuretic peptide exposure or acute exposure to phorbol esters or calcium-elevating agents causes receptor dephosphorylation and inactivation (10).…”

Section: Natriuretic Peptides and Atp Activate And Gö6976 Inhibits Gumentioning

confidence: 99%

ATP Potentiates Competitive Inhibition of Guanylyl Cyclase A and B by the Staurosporine Analog, Gö6976

Robinson

Potter

2011

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

Natriuretic peptides and ATP activate and Gö6976 inhibits guanylyl cyclase (GC)-A and GC-B. Here, the mechanism of inhibition was determined. Gö6976 progressively increased the Michaelis-Menten constant and decreased the Hill coefficient without reducing the maximal velocity of GC-A and GC-B. In the presence of 1 mM ATP, the K i was 1 M for both enzymes. Inhibition of GC-B was minimal in the absence of ATP, and 1 mM ATP increased the inhibition 4-fold. In a reciprocal manner, 10 M Gö6976 increased the potency of ATP for GC-B 4-fold. In contrast to a recent study (Duda, T., Yadav, P., and Sharma, R. K. (2010) FEBS J. 277, 2550 -2553), neither staurosporine nor Gö6976 activated GC-A or GC-B. This is the first study to show that Gö6976 reduces GTP binding and the first demonstration of a competitive inhibitor of a receptor guanylyl cyclase. We conclude that Gö6976 reduces GTP binding to the catalytic site of GC-A and GC-B and that ATP increases the magnitude of the inhibition.

show abstract

“…Furthermore, data generated from these aforementioned systems are limited in terms of reproducibility, precision and accuracy in addition to appropriate comparison with the traditional RAST. The disadvantages associated with these assays prompted us to develop a stable cell-based assay for the potency determination of therapeutic rhBNP, based on studies on GC-A receptor using HEK293 cells [32] . There are several advantages in our assay: 1) a single high-responsiveness clone was isolated with stability confirmed even a in culture for 65 passages; 2) the operation is drastically simplified with detection of cGMP in culture supernatants and use of non-radioactive materials in immunoassays in a high through-put manner; 3) the new assay has demonstrated defined specificity, greater precision and reproducibility in validation studies compared with RAST, for which no previous studies have reported.…”

Section: Discussionmentioning

confidence: 99%

A Novel Bioassay for the Activity Determination of Therapeutic Human Brain Natriuretic Peptide (BNP)

Chun-ming

Shi

et al. 2012

PLoS ONE

View full text Add to dashboard Cite

BackgroundRecombinant human brain natriuretic peptide (rhBNP) is an important peptide-based therapeutic drug indicated for the treatment of acute heart failure. Accurate determination of the potency of therapeutic rhBNP is crucial for the safety and efficacy of the drug. The current bioassay involves use of rabbit aortic strips, with experiments being complicated and time-consuming and markedly variable in results. Animal-less methods with better precision and accuracy should be explored. We have therefore developed an alternative cell-based assay, which relies on the ability of BNP to induce cGMP production in HEK293 cells expressing BNP receptor guanylyl cyclase-A.Methodology/Principal FindingsAn alternative assay based on the measurement of BNP-induced cGMP production was developed. Specifically, the bioassay employs cells engineered to express BNP receptor guanylyl cyclase-A (GCA). Upon rhBNP stimulation, the levels of the second messager cGMP in these cells drastically increased and subsequently secreted into culture supernatants. The quantity of cGMP, which corresponds to the rhBNP activity, was determined using a competitive ELISA developed by us. Compared with the traditional assay, the novel cell-based assay demonstrated better reproducibility and precision.Conclusion/SignificanceThe optimized cell-based assay is much simpler, more rapid and precise compared with the traditional assay using animal tissues. To our knowledge, this is the first report on a novel and viable alternative assay for rhBNP potency analysis.

show abstract

Arg13 of B-Type Natriuretic Peptide Reciprocally Modulates Binding to Guanylyl Cyclase but Not Clearance Receptors

Cited by 10 publications

References 14 publications

Synthesis, secretion, function, metabolism and application of natriuretic peptides in heart failure

Synthesis, secretion, function, metabolism and application of natriuretic peptides in heart failure

ATP Potentiates Competitive Inhibition of Guanylyl Cyclase A and B by the Staurosporine Analog, Gö6976

A Novel Bioassay for the Activity Determination of Therapeutic Human Brain Natriuretic Peptide (BNP)

Contact Info

Product

Resources

About