PMID: 8612617Mar 1, 1996Paper

Mutations of Glu92 in ferredoxin I from spinach leaves produce proteins fully functional in electron transfer but less efficient in supporting NADP+ photoreduction

European Journal of Biochemistry
L PiubelliG Zanetti

Abstract

Ferredoxin I in spinach chloroplasts fulfils the role of distributing electrons of low redox potential produced by photosystem I to several metabolic routes, NADP+ reduction being the major output. To investigate the role of Glu92, which is conserved in the chloroplast-type ferredoxins, mutations of this residue to either Gln, Ala or Lys were obtained through site-directed mutagenesis. A Glu93Ala mutant was also designed. The four mutants of ferredoxin I were overproduced in Escherichia coli, purified and characterised. The different migration in nondenaturing gel electrophoresis of wild-type and mutant proteins confirmed that the desired mutation was present in the expressed proteins. Spectral and physical properties of the mutants were similar to those of wild-type ferredoxin; electron-transfer properties were, however, quite different in the case of the mutants at position 92. Unexpectedly, these mutant ferredoxins were found to be twice as active as the wild-type protein in supporting the NADPH--cytochrome c reductase reaction catalysed by ferredoxin--NADP+ reductase. However, interactions of the mutant ferredoxins with the isolated thylakoid membranes deprived of endogenous ferredoxin showed that the mutants were less capa...Continue Reading

References

Dec 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·F SangerA R Coulson
Jan 22, 1991·Biochimica Et Biophysica Acta·D B Knaff, M Hirasawa
Feb 11, 1988·Nucleic Acids Research·J R SayersF Eckstein
Jun 1, 1995·Protein Expression and Purification·L PiubelliG Zanetti
Feb 1, 1994·Journal of Bioenergetics and Biomembranes·H M HoldenJ L Markley

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Citations

Jun 7, 2006·Photosynthesis Research·Giorgio FortiGiuseppe Zucchelli
Oct 24, 1998·Progress in Biophysics and Molecular Biology·H Sticht, P Rösch
Oct 24, 1998·Biochimica Et Biophysica Acta·M HipplerJ D Rochaix
Nov 20, 1997·Biochimica Et Biophysica Acta·A AlivertiG Zanetti

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