Occurrence of a para-nitrophenyl phosphate-phosphatase with calcineurin-like characteristics in Paramecium tetraurelia

Kissmehl, Roland; Treptau, Tilman; Kottwitz, Brunhilde; Plattner, Helmut

Occurrence of a para-nitrophenyl phosphate-phosphatase with calcineurin-like characteristics in Paramecium tetraurelia

Dateien

Occurrence_of_a_para.pdf(397.93 KB)

Datum

1997

Autor:innen

Kissmehl, Roland

Treptau, Tilman

Kottwitz, Brunhilde

Plattner, Helmut

Publikationstyp

Zeitschriftenartikel

Erschienen in

Archives of Biochemistry and Biophysics ; 344 (1997), 2. - S. 260-270. - ISSN 0003-9861

Zusammenfassung

Using para-nitrophenyl phosphate (pNPP) as a substrate for enzymatic activity, we sought to identify CaN inParamecium.We isolated three different pNPP-phosphatases from the soluble fraction of Parameciumcells by anion-exchange and affinity column chromatographies. One, pNPP-phosphatasePeak I, is very similar to mammalian CaN. Divalent cation dependency, inhibition by calmodulin (CaM) antagonists (trifluoperazine, calmidazolium), and insensitivity to various phosphatase inhibitors (heparin, okadaic acid, sodium vanadate, etc.) show similarity to mammalian CaN rather than to any other Paramecium pNPP-hydrolyzing enzymes tested. Polyclonal antibodies against bovine brain CaN recognizing subunits A (61 or 58 kDa) and B (17 kDa) of brain CaN cross-reacted with a 63-kDa protein in fractions containing Peak I pNPP-phosphatase activity and coeluted calmodulin. Overlay assays using biotinylated brain calmodulin indicated Ca2+-dependent CaM-binding by the 63-kDa protein. A Ca2+-binding protein with the same electrophoretic mobility as CaN B (17 kDa) was also present, though in other fractions from DEAE cellulose chromatography. This finding strongly suggests that, in the absence of Ca2+, both subunits, A and B, were separated either before or during chromatographic processing. Our data support the existence of both subunits of a CaN-like phosphatase in Paramecium cells.

Fachgebiet (DDC)

570 Biowissenschaften, Biologie

Schlagwörter

calcineurin,calmodulin,Paramecium,para-nitrophenyl phosphate,protein phosphatase

Zitieren

ISO 690

KISSMEHL, Roland, Tilman TREPTAU, Brunhilde KOTTWITZ, Helmut PLATTNER, 1997. Occurrence of a para-nitrophenyl phosphate-phosphatase with calcineurin-like characteristics in Paramecium tetraurelia. In: Archives of Biochemistry and Biophysics. 344(2), pp. 260-270. ISSN 0003-9861. Available under: doi: 10.1006/abbi.1997.0208

BibTex

@article{Kissmehl1997Occur-8039,
  year={1997},
  doi={10.1006/abbi.1997.0208},
  title={Occurrence of a para-nitrophenyl phosphate-phosphatase with calcineurin-like characteristics in Paramecium tetraurelia},
  number={2},
  volume={344},
  issn={0003-9861},
  journal={Archives of Biochemistry and Biophysics},
  pages={260--270},
  author={Kissmehl, Roland and Treptau, Tilman and Kottwitz, Brunhilde and Plattner, Helmut}
}

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