Lim, Eng-Kiat and Roberts, Michael and Bowles, D. J. (1999) Biochemical characterization of tomato annexin p35:independence of calcium-binding and phosphatase activities. Journal of Biological Chemistry, 273 (52). 34920–34925. ISSN 1083-351XFull text not available from this repository.
Tomato annexin p35 has been cloned and used in a site-directed mutagenesis study to explore the phospholipid binding and catalytic properties of the protein in detail. Analysis of the cDNA sequence of p35 reveals that the annexin has only two typical endonexin folds, corresponding to repeats I and IV. Expression of recombinant p35 in Escherichia coli confirmed both phospholipid binding and a nucleotide phosphatase activity that could be inhibited on interaction of the recombinant annexin with phospholipids. Site-directed mutagenesis in which the acidic residues Glu-68 (repeat I), and Asp-297 (repeat IV) were changed to Asn, generated two mutant forms, E68N and D297N, respectively. Both mutant forms of the annexin continued to express catalytic activity. Changing repeat I had little effect on phospholipid binding, whereas the change to repeat IV abolished this property. These data show that, in this plant annexin, repeat IV plays a more critical role in calcium-dependent phospholipid binding than repeat I, and that the catalytic and phospholipid binding activity of the protein can be separated experimentally.
|Journal or Publication Title:||Journal of Biological Chemistry|
|Uncontrolled Keywords:||annexin ; calcium ; phosphatase|
|Departments:||Faculty of Science and Technology > Lancaster Environment Centre|
|Deposited On:||20 Sep 2011 09:36|
|Last Modified:||24 Jun 2016 01:36|
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