Phosphate uptake in the yeast Candida tropicalis: purification of phosphate-binding protein and investigations about its role in phosphate uptake |
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Authors: | Robert Jeanjean Sylvie Bedu José Rocca-Serra Christian Foucault |
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Institution: | (1) Faculté des Sciences de Luminy, Laboratoire de Physiologie Cellulaire, ERA 619, case 901, 70 Route Leon Lachamp, F-13288 Marseille Cedex 9, France;(2) Faculté des Sciences de Luminy, Centre d'Immunologie, case 906, 70 Route Leon Lachamp, F-13288 Marseille Cedex 9, France |
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Abstract: | The purification of a phosphate-binding protein (PiBP2) by immunoadsorption is described. The entire anti phosphate-binding protein 2 antibodies as well as the Fab fragments obtained from these antibodies inhibit Pi uptake by whole cells. The inhibition is a mixed type of inhibition (V
m and K
m are affected). These results should be regarded as a possible involvement of phosphate-binding protein 2 in Pi uptake. The binding of 125I-labelled fragments prepared from anti phosphate-binding protein 2 antibodies to whole cells, to shocked cells and to protoplasts has been investigated. The results confirm the release of phosphate-binding protein by osmotic shock and during protoplast formation. From these findings, a cell-wall localisation, near the cell surface of the phosphate-binding protein should be proposed.Abbreviations Pi
inorganic phosphate
- PiBP
phosphate-binding protein
- Tris
Tris (hydroxymethyl)-aminoethane
- MES
(2(N-Morpholino) ethanesulfonic acid
- BSA
bovine serum albumin
- EDTA
ethylene diamine tetraacetic acid, disodium salt
- PMSF
phenylmethyl sulfonyl fluoride
- SDS
sodium dodecyl sulfate
- Fab
fragments, fragment antigen binding |
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Keywords: | Yeast Phosphate uptake Phosphate-binding protein Anti-phosphate-binding protein antibodies |
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