Localization of angiotensin converting enzyme (kininase II). I. Preparation of antibody-heme-octapeptide conjugates

J. W. Ryan, A. R. Day, D. R. Schultz, Una S. Ryan, A. Chung, D. I. Marlborough, F. E. Dorer

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Abstract

Antibodies to pig lung angiotensin converting enzyme (kininase II) were conjugated to a heme-octapeptide (8-microperoxidase, 8-MP) derived from cytochrome c. 8-MP, which has only one reactive amine, was coupled to antibody in a two-step procedure using a bifunctional active ester, bis-succinyl succinate. In the first-step, 8-MP was reacted with an excess of bis-succinyl succinate to yield 8-MP-succinyl succinate, a stable compound which can be stored. In a second step, the remaining active ester was used for coupling to reactive amines of the antibody. The conjugate consists of 1.6-2.3 8-MP moieties per antibody. Using these procedures, the formation of complex polymers is avoided. Each molecule of conjugate possesses both immuno-reactivity and peroxidatic activity. The conjugate has been used to localize angiotensin converting enzyme along the plasma membrane and associated caveolae of pig aortic endothelial cells in culture.

Original languageEnglish (US)
Pages (from-to)111-124
Number of pages14
JournalTissue and Cell
Volume8
Issue number1
DOIs
StatePublished - 1976

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ASJC Scopus subject areas

  • Developmental Biology
  • Cell Biology

Cite this

Ryan, J. W., Day, A. R., Schultz, D. R., Ryan, U. S., Chung, A., Marlborough, D. I., & Dorer, F. E. (1976). Localization of angiotensin converting enzyme (kininase II). I. Preparation of antibody-heme-octapeptide conjugates. Tissue and Cell, 8(1), 111-124. https://doi.org/10.1016/0040-8166(76)90024-0