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Purification and Properties of a Ribonuclease from Cowpea Cotyledons

Abstract

The isolation and characterisation of cotyledonary ribonucleases (RNase; EC 3.1.27.1), are basic steps to understand the physiology and biochemistry of RNA turnover and mobilisation during seed germination and seedling establishment, as well as how environmental stresses affect them. RNase was isolated and purified 928-fold, to apparent electrophoretic homogeneity from 5-d-old seedlings of Vigna unguiculata. It is a protein with an apparent molecular mass of 16 kDa having three major isoforms. Its optimum pH is 5.8, which decreases to 5.2 in presence of KCl. It has an apparent Km of 0.80 mg RNA cm-3 and retains 40 % of its activity when heated to 80 °C. It is completely inhibited by Cu2+, Hg2+ and Zn2+ and is almost insensitive to Mg2+, Ca2+- and EDTA. Urea, Fe2+, Co2+ and 2-mercaptoethanol partially inhibit its activity. Its amino acid composition shows a resem lance to that of other plant RNases.

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Gomes-Filho, E., Lima, C., Enéas-Filho, J. et al. Purification and Properties of a Ribonuclease from Cowpea Cotyledons. Biologia Plantarum 42, 525–532 (1999). https://doi.org/10.1023/A:1002602712392

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  • enzyme purification and characterization
  • RNA
  • Vigna unguiculata