Molecular isolation and characterization of novel four subisoforms of ECE-2

Ikeda, Shoko; Emoto, Noriaki; Alimsardjono, Haryanto; Yokoyama, Mitsuhiro; Matsuo, Masafumi

doi:10.1016/s0006-291x(02)00252-8

Cited by 22 publications

(14 citation statements)

References 24 publications

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“…Further- more, NL1 and angiotensin-converting enzyme have overlapping substrate specificities and hydrolyze several common peptides in vitro. Other peptidases of the M13 family of metallopeptidases such as Phex, ECE-1, ECE-2, and ECEL1 have also been detected in the testis (26,30,37,60), but their role in male fertility has not yet been established. The production of Nl1 null mice constitutes an important step in the understanding of the physiological importance of NL1 in mammals.…”

Section: Discussionmentioning

confidence: 99%

Reduced Fertility in Male Mice Deficient in the Zinc Metallopeptidase NL1

Carpentier

Guillemette

Bailey

et al. 2004

Molecular and Cellular Biology

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Members of the M13 family of zinc metalloendopeptidases have been shown to play critical roles in the metabolism of various neuropeptides and peptide hormones, and they have been identified as important therapeutic targets. Recently, a mouse NL1 protein, a novel member of the family, was identified and shown to be expressed mainly in the testis as a secreted protein. To define its physiological role(s), we used a gene targeting strategy to disrupt the endogenous murine Nl1 gene by homologous recombination and generate Nl1 mutant mice. The Nl1 ؊/؊ mice were viable and developed normally, suggesting that zygotic expression of Nl1 is not required for development. However, Nl1؊/؊ males produced smaller litters than their wild-type siblings, indicating specific male fertility problems. Reduced fertility may be explained by two impaired processes, decreased egg fertilization and perturbed early development of fertilized eggs. These two phenotypes did not result from gross anatomical modifications of the testis or from impaired spermatogenesis. Basic sperm parameters were also normal. Thus, our findings suggest that one of the roles of NL1 in mice is related to sperm function and that NL1 modulates the processes of fertilization and early embryonic development in vivo.A wide variety of biologically active peptide hormones, neuropeptides, and regulatory peptides are proteolytically activated or inactivated by zinc metalloproteases (58, 59). The M13 family of zinc metalloproteases attracts much interest because its members are involved in the metabolism of several regulatory peptides of the mammalian nervous, cardiovascular, inflammatory, and immune systems as well as in the mineralization process (8,56,57,59). For these reasons, several members of this family of peptidases have been selected as targets for therapeutic intervention by inhibitors or pharmacological agents.NL1 (also called SEP or NEP2) is the newest member of the M13 family of zinc metallopeptidases (19,25,42), which also includes neprilysin (neutral endopeptidase 24.11; also called NEP and CALLA) (reviewed in reference 11), the endothelinconverting enzymes 1 and 2 (ECE-1 and ECE-2) (reviewed in reference 55), PHEX (formerly PEX; a phosphate regulating gene with homologies to endopeptidases on the X chromosome) (reviewed in reference 54), the Kell blood group protein (reviewed in reference 35), and ECEL1/DINE (endothelinconverting enzyme-like 1/damage-induced neuronal endopeptidase) (29, 60, 61). Nl1 cDNA was simultaneously cloned by reverse transcription-PCR technology with degenerate oligonucleotide primers based on conserved sequence similarity between members of the family and mRNA isolated from mouse and rat testis or from Ece-1 Ϫ/Ϫ embryos (19,25,42). Finally, a clone coding for a putative human homologue has been obtained from a central nervous system cDNA library (4).As observed for other members of the M13 family of peptidases, the Nl1 cDNA encodes a type II transmembrane glycoprotein encompassing a short N-terminal cytoplasmic tail, a single tran...

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Section: Discussionmentioning

confidence: 99%

Reduced Fertility in Male Mice Deficient in the Zinc Metallopeptidase NL1

Carpentier

Guillemette

Bailey

et al. 2004

Molecular and Cellular Biology

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“…ECE-2 variants 2 and 5 have shown to be preferentially expressed in brain (22). Variants 2 and 5 were used for transfection of CHO cells.…”

Section: Methodsmentioning

confidence: 99%

Endothelin-converting Enzymes Degrade Intracellular β-Amyloid Produced within the Endosomal/Lysosomal Pathway and Autophagosomes*

Pacheco‐Quinto

Eckman

2013

Journal of Biological Chemistry

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Background: Endothelin-converting enzymes (ECEs) degrade ␤-amyloid (A␤) peptide. Results: ECE inhibition produces, in addition to extracellular A␤ accumulation, intracellular A␤ accumulation within endosomal/lysosomal and autophagic vesicles. Conclusion: An intracellular pool of A␤ is regulated by ECE activity at the sites of production. Significance: ECE dysfunction may cause intraneuronal A␤ accumulation, which is associated with neurotoxicity early in AD progression.

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“…In support of this, big-ET is the main species secreted from cells that co-express pre-proendothelin and an intracellular isoform of ECE-1, thus indicating the low efficiency of intracellular processing of big-ET by ECE-1 (14). On the other hand, ECE-2 could specifically fulfill this role as it displays the acidic optimum pH required for efficient conversion of precursors in the lumen of the TGN and the secretory vesicles (24), and as 2 of the 4 recently identified isoforms of ECE-2 were found in intracellular compartments (33).…”

Section: Fig 8 Ece-1 Forms Heterodimers In Double Transfectedmentioning

confidence: 99%

“…29 and 30; for review see Ref. 31), and a related gene organization has recently been described for ECE-2 (32,33). ECE-1 isoforms result from the use of alternate promoters located upstream of specific exons (29,30,34).…”

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confidence: 99%

Heterodimerization of Endothelin-converting Enzyme-1 Isoforms Regulates the Subcellular Distribution of This Metalloprotease

Muller

Barret

Etienne

et al. 2003

Journal of Biological Chemistry

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Endothelin-converting enzyme (ECE) is a membrane metalloprotease that generates endothelin from its direct precursor big endothelin. Four isoforms of ECE-1 are produced from a single gene through the use of alternate promoters. These isoforms share the same extracellular catalytic domain and contain unique cytosolic tails, which results in their specific subcellular targeting. We investigated the distribution of ECE-1 isoforms in transfected AtT-20 neuroendocrine cells. Whereas ECE-1a and 1c were present at the plasma membrane, ECE-1b and ECE-1d were retained inside the cells. We found that both intracellular isoforms were concentrated in the endosomal system: ECE-1d in recycling endosomes, and ECE-1b in late endosomes/multivesicular bodies. Leucine-based motifs were involved in the intracellular retention of these isoforms, and the targeting of ECE-1b to the degradation pathway required an additional signal in the N terminus. The concentration of ECE-1 isoforms in the endosomal system suggested new functions for these enzymes. Potential novel functions include redistribution of other isoforms through direct interaction. We have showed that ECE-1 isoforms could heterodimerize, and that in such heterodimers the ECE-1b targeting signal was dominant. Interaction of a plasma membrane isoform with ECE-1b resulted in its intracellular localization and decreased its extracellular activity. These data demonstrated that the targeting signals specific for ECE-1b constitute a regulatory domain per se that could modulate the localization and the activity of other isoforms.Endothelins (ET 1 -1, ET-2, and ET-3) are 21-residue peptides derived from three distinct genes (1). They are pleiotropic factors that play an important role in the regulation of the cardiovascular and endocrine systems (for review see Refs. 2 and 3). Endothelins are also crucial developmental factors as demonstrated by the targeted disruption of the genes coding for the precursors of ET-1 and ET-3, and of the genes coding for their receptors, ETA and ETB (4 -7). In addition, the expression of endothelin is associated with many pathological processes and with tumor growth (2, 8 -10). In order to fulfill such a wide spectrum of physiological functions, endothelins act through autocrine and paracrine mechanisms. Their biosynthesis thus requires tight local control. Endothelins are synthesized in the endoplasmic reticulum as precursors that undergo a two-step proteolytic maturation. Pro-endothelins are first processed at conserved multibasic sites by furin or a furin-like enzyme, in order to release an intermediate called big endothelin (big-ET) (11, 12), which is devoid of biological activity (13). Big-ET is then processed by endothelin converting enzyme (ECE) at a Trp-Val/Ile bond, which releases the biologically active peptide. This latter proteolytic step can occur in the extracellular medium and in the secretory pathway, so that cells secrete either big-ETs alone or together with endothelins (14, 15, 16). Endothelial cells co-express the precursor, the c...

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Molecular isolation and characterization of novel four subisoforms of ECE-2

Cited by 22 publications

References 24 publications

Reduced Fertility in Male Mice Deficient in the Zinc Metallopeptidase NL1

Reduced Fertility in Male Mice Deficient in the Zinc Metallopeptidase NL1

Endothelin-converting Enzymes Degrade Intracellular β-Amyloid Produced within the Endosomal/Lysosomal Pathway and Autophagosomes*

Heterodimerization of Endothelin-converting Enzyme-1 Isoforms Regulates the Subcellular Distribution of This Metalloprotease

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