Acid Prohormone Sequence Determines Size, Shape, and Docking of Secretory Vesicles in Atrial Myocytes

Baertschi, Alex J.; Monnier, D.; Schmidt, Frank; Levitan, Edwin S.; Fakan, Stanislas; Roatti, Angela

doi:10.1161/hh1501.095715

Cited by 50 publications

(70 citation statements)

References 23 publications

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“…3A and 6A. The pro-ANP-EGFP construct has been described elsewhere (25). Constructs were verified by automated sequencing at Eurofins Genomics (Bangalore, Karnataka, India).…”

Section: Methodsmentioning

confidence: 99%

Cytosolic Dynamics of Annexin A6 Trigger Feedback Regulation of Hypertrophy via Atrial Natriuretic Peptide in Cardiomyocytes

Banerjee¹,

Bandyopadhyay²

2014

Journal of Biological Chemistry

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Background: Altered annexin A6 expression is associated with several cardiovascular disorders, including myocyte hypertrophy, but precise functions of the protein remain elusive. Results: Dynamic association of annexin A6 with intracellular atrial natriuretic peptide precursor protects against adrenergic stimulation-induced hypertrophy of H9c2 cardiomyocytes. Conclusion: Annexin A6 negatively regulates hypertrophic progression of cardiomyocytes. Significance: Annexin A6 exhibits potential for antihypertrophic therapeutics.

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“…3A and 6A. The pro-ANP-EGFP construct has been described elsewhere (25). Constructs were verified by automated sequencing at Eurofins Genomics (Bangalore, Karnataka, India).…”

Section: Methodsmentioning

confidence: 99%

Cytosolic Dynamics of Annexin A6 Trigger Feedback Regulation of Hypertrophy via Atrial Natriuretic Peptide in Cardiomyocytes

Banerjee¹,

Bandyopadhyay²

2014

Journal of Biological Chemistry

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“…Detection of a subnormal status of these B vitamins has considerable importance. Vitamin B 12 deficiency in the elderly may cause cognitive dysfunction (5 ), neuropsychiatric disorders, dementia (6 ), and hyperhomocysteinemia (5 ), an independent risk factor for coronary vascular disease and dementia (7,8 ). A 5 mol/L increment in total plasma homocysteine (tHcy) has been associated with a 49% increase in all-cause of mortality among individuals 65-72 years of age (9 ).…”

Section: Quantification Of 56-dihydrouracil By Hplc-electromentioning

confidence: 99%

Quantification of 5,6-Dihydrouracil by HPLC–Electrospray Tandem Mass Spectrometry

et al. 2004

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“…However, under certain conditions, the N-terminal region might be minimally accessible for antibody binding (7,8 ). Despite the described long half-life of proANP, results from various competitive immunoassays as well as HPLC analyses indicate that proANP1-98 can be subject to further fragmentation (9, 10 ).…”

mentioning

confidence: 99%

Immunoluminometric Assay for the Midregion of Pro-Atrial Natriuretic Peptide in Human Plasma

Morgenthaler¹,

Struck²,

Thomas³

et al. 2004

Clinical Chemistry

316

285

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The prohormone of atrial natriuretic peptide (proANP) is a polypeptide of 126 amino acids. Mature atrial natriuretic peptide (ANP) consists of amino acids 99 -126 and comprises 98% of the natriuretic peptides in the circulation (1 ). The N-terminal portion of proANP, termed proANP1-98 or NT-proANP, has a much longer half-life than mature ANP and has therefore been suggested to be a more reliable analyte for measurement than mature ANP (2 ). In addition to other established indications, proANP has recently gained much interest as a potential new marker in the field of sepsis (3 ), emphasizing the need for a reliable assay for this molecule.All sandwich immunoassays developed for proANP to date use an antibody against the N-terminal region of proANP1-98 combined with a second antibody against either the midregion (4 ) or C-terminal region (5, 6 ). However, under certain conditions, the N-terminal region might be minimally accessible for antibody binding (7,8 ). Despite the described long half-life of proANP, results from various competitive immunoassays as well as HPLC analyses indicate that proANP1-98 can be subject to further fragmentation (9, 10 ).We developed a new sandwich immunoassay for midregional proANP (amino acids 53-90; EDTA-plasma samples were collected at a blood bank (Red Cross) from 325 consecutive healthy blood donors (age range, 18 -67 years; 52.9% male) without clinical evidence of acute disease or a history of chronic illness. Blood donors with risk factors for heart failure were not enrolled in the study. Written consent was obtained from all donors.For the proANP assay, tubes were coated with affinitypurified polyclonal sheep antibodies specific for amino acids 73-90 (GRGPWDSSDRSALLKSKL) of the molecule (Fig. 1A). Coating of the antibody was done for 20 h on polystyrene tubes (1.5 g/tube) in 0.3 mL of buffer (10 mmol/L Tris-HCl, pH 7.8; 10 mmol/L NaCl). Tubes were blocked with 10 mmol/L sodium phosphate buffer containing 30 g/L of the polysaccharide Karion FP (Merck AG) and 5 g/L protease-free bovine serum albumin (Sigma), pH 6.8, and were lyophilized. A polyclonal sheep antibody specific for another part of proANP was used as tracer. This antibody was raised to peptide 53-72 (PEVP-PWTGEVSPAQRDGGAL) of proANP and was affinity purified on a peptide-sulfolink column. After purification, the antibody was labeled with acridinium ester as follows: 100 g of antibody in 20 mmol/L sodium phosphate buffer, pH 8.0, was incubated for 20 min at room temperature with 10 L of acridinium ester (1 g/L in acetonitrile; Hoechst AG). Labeled antibody was purified by HPLC with a Knauer hydroxyapatite column (buffer gradient, 1-500 mmol/L potassium phosphate, pH 6.8; flow rate, 0.8 mL/min).Plasma proANP was measured as follows: 20 L of patient sample (EDTA plasma) or calibrator was added in duplicate to antibody-coated tubes containing 50 mmol/L sodium phosphate buffer, pH 7.5, and incubated for 2 h at room temperature. After five washes with 2 mL of standard LUMItest © washing buffer (B.R.A.H.M.S AG), 200 L of t...

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Acid Prohormone Sequence Determines Size, Shape, and Docking of Secretory Vesicles in Atrial Myocytes

Abstract: Abstract-How vesicles are born in the trans-Golgi network and reach their docking sites at the plasma membrane is still largely unknown and is investigated in the present study on live, primary cultured atrial cardiomyocytes.

Cited by 50 publications

References 23 publications

Cytosolic Dynamics of Annexin A6 Trigger Feedback Regulation of Hypertrophy via Atrial Natriuretic Peptide in Cardiomyocytes

Cytosolic Dynamics of Annexin A6 Trigger Feedback Regulation of Hypertrophy via Atrial Natriuretic Peptide in Cardiomyocytes

Quantification of 5,6-Dihydrouracil by HPLC–Electrospray Tandem Mass Spectrometry

Immunoluminometric Assay for the Midregion of Pro-Atrial Natriuretic Peptide in Human Plasma

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