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Biochemical characterization of tomato annexin p35: independence of calcium-binding and phosphatase activities

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Biochemical characterization of tomato annexin p35: independence of calcium-binding and phosphatase activities. / Lim, Eng-Kiat; Roberts, Michael; Bowles, D. J.
In: Journal of Biological Chemistry, Vol. 273, No. 52, 25.12.1999, p. 34920–34925.

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Lim E-K, Roberts M, Bowles DJ. Biochemical characterization of tomato annexin p35: independence of calcium-binding and phosphatase activities. Journal of Biological Chemistry. 1999 Dec 25;273(52):34920–34925. doi: 10.1074/jbc.273.52.34920

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Lim, Eng-Kiat ; Roberts, Michael ; Bowles, D. J. / Biochemical characterization of tomato annexin p35 : independence of calcium-binding and phosphatase activities. In: Journal of Biological Chemistry. 1999 ; Vol. 273, No. 52. pp. 34920–34925.

Bibtex

@article{23e20d5391af4a008424ac09be5b3d77,
title = "Biochemical characterization of tomato annexin p35: independence of calcium-binding and phosphatase activities",
abstract = "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 mutantforms 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.",
keywords = "annexin, calcium, phosphatase",
author = "Eng-Kiat Lim and Michael Roberts and Bowles, {D. J.}",
year = "1999",
month = dec,
day = "25",
doi = "10.1074/jbc.273.52.34920",
language = "English",
volume = "273",
pages = "34920–34925",
journal = "Journal of Biological Chemistry",
issn = "1083-351X",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "52",

}

RIS

TY - JOUR

T1 - Biochemical characterization of tomato annexin p35

T2 - independence of calcium-binding and phosphatase activities

AU - Lim, Eng-Kiat

AU - Roberts, Michael

AU - Bowles, D. J.

PY - 1999/12/25

Y1 - 1999/12/25

N2 - 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 mutantforms 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.

AB - 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 mutantforms 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.

KW - annexin

KW - calcium

KW - phosphatase

U2 - 10.1074/jbc.273.52.34920

DO - 10.1074/jbc.273.52.34920

M3 - Journal article

VL - 273

SP - 34920

EP - 34925

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 1083-351X

IS - 52

ER -