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Homologous substitution of ACE C-domain regions with N-domain sequences: effect on processing, shedding, and catalytic properties

  • Zenda L. Woodman , Sylva L.U. Schwager , Pierre Redelinghuys , Anthony J. Chubb , Elizabeth L. van der Merwe , Mario R.W. Ehlers and Edward D. Sturrock
Published/Copyright: August 9, 2006
Biological Chemistry
From the journal Volume 387 Issue 8

Abstract

Angiotensin-converting enzyme (ACE) exists as two isoforms: somatic ACE (sACE), comprised of two homologous N and C domains, and testis ACE (tACE), comprised of the C domain only. The N and C domains are both active, but show differences in substrate and inhibitor specificity. While both isoforms are shed from the cell surface via a sheddase-mediated cleavage, tACE is shed much more efficiently than sACE. To delineate the regions of tACE that are important in catalytic activity, intracellular processing, and regulated ectodomain shedding, regions of the tACE sequence were replaced with the corresponding N-domain sequence. The resultant chimeras C1–163Ndom-ACE, C417–579Ndom-ACE, and C583–623Ndom-ACE were processed to the cell surface of transfected Chinese hamster ovary (CHO) cells, and were cleaved at the identical site as that of tACE. They also showed acquisition of N-domain-like catalytic properties. Homology modelling of the chimeric proteins revealed structural changes in regions required for tACE-specific catalytic activity. In contrast, C164–416Ndom-ACE and C191–214Ndom-ACE demonstrated defective intracellular processing and were neither enzymatically active nor shed. Therefore, critical elements within region D164–V416 and more specifically I191–T214 are required for the processing, cell-surface targeting, and enzyme activity of tACE, and cannot be substituted for by the homologous N-domain sequence.

Keywords: cooperativity; domain selectivity; proteolytic cleavage; sheddase
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Published Online: 2006-08-09
Published in Print: 2006-08-01

©2006 by Walter de Gruyter Berlin New York

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https://doi.org/10.1515/BC.2006.129
Keywords for this article
cooperativity; domain selectivity; proteolytic cleavage; sheddase

Articles in the same Issue

  1. Caspase-containing complexes in the regulation of cell death and inflammation
  2. Regulation of human cathepsin B by alternative mRNA splicing: homeostasis, fatal errors and cell death
  3. The peptidases from fungi and viruses
  4. C. elegans as a model system to study the function of the COG complex in animal development
  5. Functional responses of bone cells to thrombin
  6. Homologous substitution of ACE C-domain regions with N-domain sequences: effect on processing, shedding, and catalytic properties
  7. Production and processing of a recombinant Fasciola hepatica cathepsin B-like enzyme (FhcatB1) reveals potential processing mechanisms in the parasite
  8. Development of a red-shifted fluorescence-based assay for SARS-coronavirus 3CL protease: identification of a novel class of anti-SARS agents from the tropical marine sponge Axinella corrugata
  9. Single-cell resolution imaging of membrane-anchored hepatitis C virus NS3/4A protease activity
  10. Treatment of MCF-7 cells with taxol and etoposide induces distinct alterations in the expression of apoptosis-related genes BCL2, BCL2L12, BAX, CASPASE-9 and FAS
  11. Proteolytic mechanism of a novel mitochondrial and chloroplastic PreP peptidasome
  12. Tripeptidyl-peptidase I in health and disease
  13. Molecular and functional analysis of new members of the wheat PR4 gene family
  14. C-Terminal truncations of syncytin-1 (ERVWE1 envelope) that increase its fusogenicity
  15. Disease processes may be reflected by correlations among tissue kallikrein proteases but not with proteolytic factors uPA and PAI-1 in primary ovarian carcinoma
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  17. Adaptation of the behaviour of an aspartic proteinase inhibitor by relocation of a lysine residue by one helical turn
  18. Cathepsins L and S are not required for activation of dipeptidyl peptidase I (cathepsin C) in mice
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