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      https://www.ias.ac.in/article/fulltext/jbsc/002/03/0211-0225

    • Keywords

       

      Phaseolus aureus ; nucleotide pyrophosphatase; regulation; desensitization; conformational changes; circular dichroism; optical rotatory dispersion

    • Abstract

       

      A homogenous and crystalline form of nucleotide pyrophosphatase (EC 3.6.1.9) fromPhaseolus aureus (mung bean) seedlings was used for the study of the regulation of enzyme activity by adenine nucleotides. The native dimeric form of the enzyme had a helical content of about 65% which was reduced to almost zero values by the addition of AMP. In addition to this change in the helical content, AMP converted the native dimer to a tetramer. Desensitization of AMP regulation, without an alteration of the molecular weight, was achieved either by reversible denaturation with 6 M urea or by passage through a column of Blue Sepharose but additionofp-hydroxymercuribenzoate desensitized the enzyme by dissociating the native dimer to a monomer. The changes in the quaternary structure and conformation of the enzyme consequent to AMP interaction or desensitization were monitored by measuring the helical content, EDTA inactivation and Zn2+ reactivation, stability towards heat denaturation, profiles of urea denaturation and susceptibility towards proteolytic digestion. Based on these results and our earlier work on this enzyme, we propose a model for the regulation of the mung bean nucleotide pyrophosphatase by association-dissociation and conformational changes. The model emphasizes that multiple mechanisms are operative in the desensitization of regulatory proteins.

    • Author Affiliations

       

      A R Venugopala Reddy1 2 C V Balakrishnan1 2 3 J Sobhanaditya1 2 S D Ravindranath1 2 4 V S Ananthanarayanan1 2 N Appaji Rao1 2

      1. Department of Biochemistry, and Molecular Biophysics Unit, Indian Institute of Science, Bangalore - 560 012
      2. Department of Biochemistry, Memorial University of Newfoundland St. John’s, Newfoundland, Canada
      3. Division of Neurology, Department of Medicine, Duke University Medical Centre, Durham, North Carolina, USA
      4. Department of Biochemistry, Toklai Experimental Station, Jorhat, Assam - 785 008, India
    • Dates

       
  • Journal of Biosciences | News

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