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Reactivity of sulphydryl groups of cytosolic and mitochondrial bovine aspartate aminotransferases

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Summary

Reactivity of sulphydryl groups of cytosolic and mitochondrial aspartate aminotransferases from ox heart has been studied. A total of 5 and 7 cysteine residues per monomer are present in cAATo and mAATo, respectively. In native conditions only a single sulphydryl group can be titrated by Nbs2 while the catalytic activity remains unchanged, however in the mitochondrial isozyme the reactivity depends on the functional state of the enzyme. Reactivity toward NEM reveals the existence of a syncatalytic sulphydryl group in the cytosolic isozyme. Titration of cAATo with pMB at pH 8 and pH 5 confirms the existence of two exposed sulphydryl groups with a different reactivity. The results compared with those reported on the corresponding isozymes from pig and chicken heart show that syncatalytic sulphydryl groups are of general occurrence in these enzymes.

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Abbreviations

cAAT:

cytosolic aspartate aminotransferase

mAAT:

mitochondrial aspartate aminotransferase

the suffixesp :

c ando stands for pig, chicken and ox heart respectively

NEM:

N-ethylmaleimide

Nbs2 :

5,5′-dithiobis(2-nitrobenzoate)

Nbs:

5-thio(2-nitrobenzoate)

SDS:

sodium dodecyl sulfate

pMB:

p-mercuribenzoate.Enzyme. Aspartate aminotransferase (EC 2.6.1.1)

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Author notes

  1. Pasquale Petrilli

    Present address: Istituto di Industrie Agrarie, Università di Napoli, Napoli, Italia

Authors and Affiliations

  1. Istituto di Chimica Organicae Biologica, Università di Napoli, Napoli, Italia

    Pasquale Petrilli, Pietro Pucci, Anna Maria Garzillo, Giovanni Sannia &  Gennaro Marino

  2. Facoltà di Scienze, Instituto di Chimica Organica e Biologica dell'Università, Via Mezzocannone, 16, 80134, Napoli, Italy

    Gennaro Marino

Authors
  1. Pasquale Petrilli
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  2. Pietro Pucci
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  4. Giovanni Sannia
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  5. Gennaro Marino
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Petrilli, P., Pucci, P., Garzillo, A.M. et al. Reactivity of sulphydryl groups of cytosolic and mitochondrial bovine aspartate aminotransferases. Mol Cell Biochem 35, 121–128 (1981). https://doi.org/10.1007/BF02354826

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