Biochemical characterization of a high affinity phosphate transporter (PiPT) from root endophyte fungus Piriformospora indica.
Protein Expr Purif
; 223: 106559, 2024 Nov.
Article
em En
| MEDLINE
| ID: mdl-39089400
ABSTRACT
We have functionally characterized the high-affinity phosphate transporter (PiPT) from the root endophyte fungus Piriformospora indica. PiPT belongs to the major facilitator superfamily (MFS). PiPT protein was purified by affinity chromatography (Ni-NTA) and Size Exclusion Chromatography (SEC). The functionality of solubilized PiPT was determined in detergent-solubilized state by fluorescence quenching and in proteoliposomes. In the fluorescence quenching assay, PiPT exhibited a saturation concentration of approximately 2 µM, at a pH of 4.5. Proteoliposomes of size 121.6 nm radius, showed transportation of radioactive phosphate. Vmax was measured to be 232.2 ± 11 pmol/min/mg protein. We have found Km to be 45.8 ± 6.2 µM suggesting high affinity towards phosphate.
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01-internacional
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MEDLINE
Assunto principal:
Basidiomycota
/
Proteínas de Transporte de Fosfato
Idioma:
En
Revista:
Protein Expr Purif
Ano de publicação:
2024
Tipo de documento:
Article