Epitope mapping of mAbs to denatured human testicular ACE (CD143)

I. V. Balyasnikova, R. Metzger, F. E. Franke, N. Conrad, H. Towbin, D. E. Schwartz, E. D. Sturrock, S. M. Danilov

Research output: Contribution to journalArticlepeer-review

15 Scopus citations

Abstract

Angiotensin I-converting enzyme (ACE; CD143) has two homologous enzymatically active domains (N and C) and plays a crucial role in blood pressure regulation and vascular remodeling. A wide spectrum of monoclonal antibodies (mAbs) to different epitopes on the N and C domains of human ACE have been used to study different aspects of ACE biology. In this study, we characterized a set of nine mAbs, developed against the C domain of human ACE, which recognize the denatured forms of ACE and thus are suitable for the detection and quantification of somatic ACE (sACE) and testicular ACE (tACE) using Western blotting and immunohistochemistry on paraffin-embedded human tissues. The epitopes for these mAbs were defined using species cross-reactivity, phage display library screening, Western blotting and ACE mutagenesis. Most of the mAbs recognized common/overlapping region(s) on both somatic and testicular forms of human ACE, whereas mAb 4E10 was relatively specific for the testicular isoform and mAb 5B9 mainly recognized the glycan attached to Asn 731. This set of mAbs is useful for identifying even subtle changes in human ACE conformation because of denaturation. These mAbs are also sensitive tools for the detection of human sACE and tACE in biological fluids and tissues using proteomic approaches. Their high reactivity in paraffin-embedded tissues provides opportunities to study changes in the pattern of ACE expression and glycosylation (particularly with mAb 5B9) in different tissues and cells.

Original languageEnglish (US)
Pages (from-to)354-368
Number of pages15
JournalTissue Antigens
Volume72
Issue number4
DOIs
StatePublished - Oct 1 2008

Keywords

  • Angiotensin I-converting enzyme
  • CD143
  • Immunohistochemistry
  • Western blotting

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Biochemistry
  • Genetics

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