A mutant of spinach ferredoxin-NADP+ reductase, in which Lys-88 has been changed to glutamine, has been obtained by site-directed mutagenesis. The mutant enzyme was fully active as a diaphorase, but partially impaired in ferredoxin-dependent cytochrome c reductase activity. By steady-state kinetics, the Km for ferredoxin of the K88Q enzyme was found to have increased 10-fold, whereas the kcat was unaffected by the amino acid replacement. The interaction between oxidized ferredoxin and the enzyme forms was also studied by spectrofluorimetric titration: Kd values of 110 and 10 nM were determined for the mutant and wild-type proteins, respectively. These data point out the importance of a positive charge at position 88 of the reductase for the interaction with ferredoxin, confirming previous cross-linking studies.
Involvement of lysine-88 of spinach ferredoxin-NADP+ reductase in the interaction with ferredoxin / A. Aliverti, M.E. Corrado, G. Zanetti. - In: FEBS LETTERS. - ISSN 0014-5793. - 343:3(1994 May 02), pp. 247-250.
|Titolo:||Involvement of lysine-88 of spinach ferredoxin-NADP+ reductase in the interaction with ferredoxin|
ALIVERTI, ALESSANDRO (Primo)
ZANETTI, GIULIANA (Ultimo)
|Parole Chiave:||Electrostatic interaction; Ferredoxin; Ferredoxin-NADP+ reductase; Flavoprotein; Protein engineering; Protein-protein interaction|
|Settore Scientifico Disciplinare:||Settore BIO/10 - Biochimica|
Settore BIO/11 - Biologia Molecolare
Settore BIO/04 - Fisiologia Vegetale
|Data di pubblicazione:||2-mag-1994|
|Digital Object Identifier (DOI):||http://dx.doi.org/10.1016/0014-5793(94)80565-2|
|Appare nelle tipologie:||01 - Articolo su periodico|