Role of the Hydrophobic Transmembrane Domain in Membrane Anchoring of Endothelin-Converting Enzyme-1a

2000 ◽  
Vol 36 (Supplement 1) ◽  
pp. S30-S32
Author(s):  
Sharon Courtnie Brooks ◽  
Adviye Ergul
Keyword(s):  
Transmembrane Domain ◽  
Converting Enzyme ◽  
Endothelin Converting Enzyme ◽  
Hydrophobic Transmembrane Domain ◽  
Membrane Anchoring

Role of endothelin-converting enzyme, chymase and neutral endopeptidase in the processing of big ET-1, ET-1(1-21) and ET-1(1-31) in the trachea of allergic mice

2002 ◽  
Vol 103 (s2002) ◽  
pp. 353S-356S ◽  
Author(s):  
Benjamin A. DE CAMPO ◽  
Roy G. GOLDIE ◽  
Arco Y. JENG ◽  
Peter J. HENRY
Keyword(s):  
Mast Cell ◽  
Mast Cells ◽  
Response Curve ◽  
Histological Analysis ◽  
Neutral Endopeptidase ◽  
Converting Enzyme ◽  
Rightward Shift ◽  
Endothelin Converting Enzyme ◽  
Mast Cell Chymase

The present study examined the roles of endothelin-converting enzyme (ECE), neutral endopeptidase (NEP) and mast cell chymase as processors of the endothelin (ET) analogues ET-1(1–21), ET-1(1–31) and big ET-1 in the trachea of allergic mice. Male CBA/CaH mice were sensitized with ovalbumin (10µg) delivered intraperitoneal on days 1 and 14, and exposed to aerosolized ovalbumin on days 14, 25, 26 and 27 (OVA mice). Mice were killed and the trachea excised for histological analysis and contraction studies on day 28. Tracheae from OVA mice had 40% more mast cells than vehicle-sensitized mice (sham mice). Ovalbumin (10µg/ml) induced transient contractions (15±3% of the Cmax) in tracheae from OVA mice. The ECE inhibitor CGS35066 (10µM) inhibited contractions induced by big ET-1 (4.8-fold rightward shift of dose-response curve; P<0.05), but not those induced by either ET-1(1–21) or ET-1(1–31). The chymase inhibitors chymostatin (10µM) and Bowman-Birk inhibitor (10µM) had no effect on contractions induced by any of the ET analogues used. The NEP inhibitor CGS24592 (10µM) inhibited contractions induced by ET-1(1–31) (6.2-fold rightward shift; P<0.05) but not ET-1(1–21) or big ET-1. These data suggest that big ET-1 is processed predominantly by a CGS35066-sensitive ECE within allergic airways rather than by mast cell-derived proteases such as chymase. If endogenous ET-1(1–31) is formed within allergic airways, it is likely to undergo further conversion by NEP to more active products.

scholarly journals The Role of Endothelin-Converting Enzyme-1 in the Development of α 1 -Adrenergic-Stimulated Hypertrophy in Cultured Neonatal Rat Cardiac Myocytes

Circulation ◽  
1999 ◽  
Vol 99 (2) ◽  
pp. 292-298 ◽  
Author(s):  
Satoshi Kaburagi ◽  
Koji Hasegawa ◽  
Tatsuya Morimoto ◽  
Makoto Araki ◽  
Tatsuya Sawamura ◽  
...  
Keyword(s):  
Cardiac Myocytes ◽  
Neonatal Rat ◽  
Converting Enzyme ◽  
Endothelin Converting Enzyme ◽  
Neonatal Rat Cardiac Myocytes ◽  
Rat Cardiac Myocytes

Epitope-specific antibody-induced cleavage of angiotensin-converting enzyme from the cell surface

2002 ◽  
Vol 362 (3) ◽  
pp. 585-595 ◽  
Author(s):  
Irina V. BALYASNIKOVA ◽  
Eric H. KARRAN ◽  
Ronald F. ALBRECHT ◽  
Sergei M. DANILOV
Keyword(s):  
Transmembrane Domain ◽  
Umbilical Vein ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Integral Membrane Protein ◽  
Converting Enzyme ◽  
Ovary Cells ◽  
Angiotensin I Converting Enzyme ◽  
Hydrophobic Transmembrane Domain ◽  
Umbilical Vein Endothelial Cells

Angiotensin I-converting enzyme (ACE; CD143, EC 3.4.15.1) is a type-1 integral membrane protein that can also be released into extracellular fluids (such as plasma, and seminal and cerebrospinal fluids) as a soluble enzyme following cleavage mediated by an unidentified protease(s), referred to as ACE secretase, in a process known as ‘shedding'. The effects of monoclonal antibodies (mAbs) to eight different epitopes on the N-terminal domain of ACE on shedding was investigated using Chinese hamster ovary cells (CHO cells) expressing an ACE transgene and using human umbilical vein endothelial cells. Antibody-induced shedding of ACE was strongly epitope-specific: most of the antibodies increased the shedding by 20–40%, mAbs 9B9 and 3A5 increased the shedding by 270 and 410% respectively, whereas binding of mAb 3G8 decreased ACE shedding by 36%. The ACE released following mAb treatment lacked a hydrophobic transmembrane domain anchor. The antibody-induced shedding was completely inhibited at 4°C and by zinc chelation using 1,10-phenanthroline, suggesting involvement of a metalloprotease in this process. A hydroxamate-based metalloprotease inhibitor (batimastat, BB-94) was 15 times more efficacious in inhibiting mAb-induced ACE shedding than basal (constitutive) ACE release. Treatment of CHO-ACE cells with BB-94 more effectively prevented elevation in antibody-dependent (but not basal) ACE release induced by 3,4-dichloroisocoumarin and iodoacetamide. These data suggest that different secretases might be responsible for ACE release under basal compared with antibody-induced shedding. Further experiments with more than 40 protease inhibitors suggest that calpains, furin and the proteasome may participate in this process.

scholarly journals Endothelin-converting enzyme-1 increases in atherosclerotic mice: potential role of oxidized low density lipoproteins

2008 ◽  
Vol 50 (3) ◽  
pp. 364-375 ◽  
Author(s):  
Patricia Martínez-Miguel ◽  
Viviana Raoch ◽  
Carlos Zaragoza ◽  
Jose Manuel Valdivielso ◽  
Manuel Rodríguez-Puyol ◽  
...  
Keyword(s):  
Potential Role ◽  
Low Density Lipoproteins ◽  
Low Density ◽  
Converting Enzyme ◽  
Oxidized Low Density Lipoproteins ◽  
Endothelin Converting Enzyme

scholarly journals Role of bradykinin and endothelin-converting enzyme-1 in pulmonary hypertension

Life Sciences ◽  
2013 ◽  
Vol 93 (25-26) ◽  
pp. e33
Author(s):  
Sunu B. Raharjo ◽  
Noriaki Yuniadi ◽  
Yoga Emoto ◽  
Kazuhiko Nakayama ◽  
Ganesja M./INS; Harimurti
Keyword(s):  
Pulmonary Hypertension ◽  
Converting Enzyme ◽  
Endothelin Converting Enzyme

On the role of endothelin-converting enzyme-1 (ECE-1) and neprilysin in human breast cancer

2007 ◽  
Vol 106 (3) ◽  
pp. 361-369 ◽  
Author(s):  
Martin Smollich ◽  
Martin Götte ◽  
George W. Yip ◽  
Eng-Siang Yong ◽  
Christian Kersting ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Human Breast Cancer ◽  
Converting Enzyme ◽  
Human Breast ◽  
Endothelin Converting Enzyme

Abstract 12676: C-Terminal Residues of Phospholamban are Critical Determinants of the Quaternary Structure and Function of the Calcium ATPase Regulatory Complex

Circulation ◽  
2014 ◽  
Vol 130 (suppl_2) ◽  
Author(s):  
Neha Abrol ◽  
Nikolai Smolin ◽  
Delaine K Ceholski ◽  
Howard S Young ◽  
Seth L Robia
Keyword(s):  
Fluorescent Protein ◽  
Quaternary Structure ◽  
Transmembrane Domain ◽  
Resonance Energy ◽  
Separation Distance ◽  
Significant Loss ◽  
Donor And Acceptor ◽  
Regulatory Complex ◽  
Membrane Anchoring

Rationale: A naturally occurring missense Leu-39stop (L39X) mutation in phospholamban (PLB) results in truncation of the C-terminal transmembrane domain, leading to cardiomyopathy and premature death in humans. Objective: The goal of this study was to determine the structural and regulatory role of the C-terminal residues of PLB in the membranes of living cells. Methods and Results: We fused fluorescent protein tags to PLB and cardiac Ca 2+ ATPase (SERCA) to investigate the role of PLB C-terminal residues for membrane localization, PLB oligomerization and SERCA regulation. Alanine substitution of C-terminal residues significantly altered fluorescence resonance energy transfer (FRET) from PLB to PLB and SERCA to PLB. Notably, substitution mutation V49A had profound effects on pentamer structure and regulatory complex conformation, increasing and decreasing probe separation distance, respectively. Progressive deletion of only a few C-terminal residues resulted in significant loss of PLB membrane anchoring and mislocalization to the cytoplasm and nucleus. Selective permeabilization of the plasma membrane by saponin resulted in diffusion of fluorescently labeled PLB out of the cells, consistent with solubilization of truncated proteins. Molecular dynamics simulations recapitulated decreased bilayer anchoring for truncated PLB. C-terminal truncations resulted in progressive loss of PLB-PLB FRET, due to a decrease in the apparent affinity of PLB oligomerization. We quantified a similar decrease in the SERCA-PLB binding affinity, and loss of inhibitory potency as quantified by Ca 2+ -dependent ATPase activity. However, despite decreased SERCA-PLB binding, intermolecular FRET was paradoxically increased as a result of a 14.5 Å decrease in the distance between donor and acceptor fluorophores. Conclusions: We conclude that PLB C-terminal residues are critical for membrane anchoring and quaternary structure determination of PLB pentamer and PLB-SERCA regulatory complex. The loss of membrane registration restraint by C-terminal residues (especially V49) causes displacement of PLB to an alternative position on SERCA. The data are compatible with a model in which PLB binds to the canonical inhibitory binding site and an additional novel site.

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