biologia plantarum

International journal on Plant Life established by Bohumil Nìmec in 1959

Biologia plantarum 42:525-532, 1999 | DOI: 10.1023/A:1002602712392

Purification and Properties of a Ribonuclease from Cowpea Cotyledons

E. Gomes-Filho1, C.R.F.M. Lima1, J. Enéas-Filho1, F.A.P. Campos1, L.A. Gondim1, J.T. Prisco1
1 Department of Biochemistry and Molecular Biology, Federal University of Ceará, Fortaleza, Ceará, Brazil

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.

Keywords: enzyme purification and characterization; RNA; Vigna unguiculata
Subjects: cobalt, ribonuclease; copper, ribonuclease; cotyledon, ribonuclease purification, properties; cowpea cotyledons, RNA, RNase; enzyme purification and characterisation; heavy metals, ribonuclease; iron, Fe2+, ribonuclease; mercury, ribonuclease; ribonuclease, purification, properties; urea, ribonuclease; Vigna unguiculata; zinc, ribonuclease inhibition

Published: December 1, 1999  Show citation

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Gomes-Filho, E., Lima, C.R.F.M., Enéas-Filho, J., Campos, F.A.P., Gondim, L.A., & Prisco, J.T. (1999). Purification and Properties of a Ribonuclease from Cowpea Cotyledons. Biologia plantarum, 42(4), 525-532. doi: 10.1023/A:1002602712392
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