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A test for prorelaxin-processing enzymes using unmodified peptide substrates

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Summary

Relaxin is a member of the insulin family of proteins. It is produced principally in ovarian cells by processing of its larger precursor, prorelaxin. The enzymes responsible for conversion of prorelaxin to the mature hormone have not yet been elucidated. A rapid and convenient test has been developed to detect prorelaxin-processing enzymes in porcine ovary extracts. Unmodified peptide substrates, which represent the two prorelaxin-processing sites, were chemically synthesised and nanomolar amounts of these substrates were incubated in solution with enzyme preparations. The resultant fragments were resolved using high performance liquid chromatography or capillary electrophoresis and identified by their retention times compared with synthetic standards. This test has been successfully used to identify and characterise a candidate prorelaxin-processing enzyme from chromatographically fractionated porcine ovary extracts. The enzyme was found to be a serine protease with preference for cleavage after tetrabasic sequences and with optimal activity at pH 5.5–6.5.

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Authors and Affiliations

  1. Howard Florey Institute of Experimental Physiology and Medicine, University of Melbourne, 3052, Parkville, Victoria, Australia

    Selena S. Layden & Geoffrey W. Tregear

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  1. Selena S. Layden
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  2. Geoffrey W. Tregear
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Layden, S.S., Tregear, G.W. A test for prorelaxin-processing enzymes using unmodified peptide substrates. Lett Pept Sci 2, 83–92 (1995). https://doi.org/10.1007/BF00128502

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  • DOI: https://doi.org/10.1007/BF00128502

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