L-asparaginase of Tetrahymena Pyriformis is Associated with a Kinase Activity

Overview

Journal Mol Cell Biochem

Publisher Springer

Specialty Biochemistry

Date 1990 Jun 1

PMID 2114526

Citations 2

Authors

S A Tsirka

D A Kyriakidis

Affiliations

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Abstract

Most of L-asparaginase activity of Tetrahymena pyriformis was found to be present in microsomal membranes from which it has been purified to homogeneity (Tsirka, S.A.E. and Kyriakidis, D.A. Mol. Cell. Biochem. 83: 147-155, 1988). The native enzyme has a relative molecular weight of approximately 200 kDa, while under denaturing conditions the enzyme exhibits a subunit size of 39 kDa. Aminoacid analysis and an oligopeptide from N-terminal sequence have been determined. Dephosphorylation of L-asparaginase by alkaline phosphatase results in an activation of its catalytic activity. This enzyme also exhibits intrinsic phosphorylation activity with a Km value for ATP of 0.5 mM. Autophosphorylation with [gamma-32P] ATP of purified L-asparaginase results in the phosphorylation of tyrosine residues as well as in loss of its activity. Mg2+ and Ca2+ added together act synergistically to stimulate the kinase activity by more than 160%. The polyamines putrescine, spermidine and spermine activate the kinase approximately 100%, while neither cAMP or cGMP have any effect. These results indicate that this membrane protein with dual L-asparaginase/kinase activity must play an important role in regulating the intracellular levels of L-asparagine in Tetrahymena pyriformis.

Citing Articles

L-asparaginase of Thermus thermophilus: purification, properties and identification of essential amino acids for its catalytic activity.

Pritsa A, Kyriakidis D Mol Cell Biochem. 2001; 216(1-2):93-101.

PMID: 11216870 DOI: 10.1023/a:1011066129771.

Antiproliferative activity of L-asparaginase of Tetrahymena pyriformis on human breast cancer cell lines.

Kyriakidis D, Tsirka S, Tsavdaridis I, Iliadis S, Kortsaris A Mol Cell Biochem. 1990; 96(2):137-42.

PMID: 2125695 DOI: 10.1007/BF00420905.

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