Site-specific mutagenesis of Rhodobacter capsulatus ferredoxin I, FdxN, that functions in nitrogen fixation. Role of extra residues.

The Journal of Biological Chemistry
K SaekiS Itoh

Abstract

One of the two [4Fe-4S]-type clusters of the Rhodobacter capsulatus ferredoxin I, FdxN, was modified through site-specific mutagenesis of the distinctive features of the second cluster-binding motif, Cys38-X2-Cys41-X8-Cys50-X3-Cys54-X4-Cys59. First, various mutagenized products were tested to learn whether they could rescue the decreased capacity of an fdxN-null strain MSA1 to fix nitrogen: the phenotype of MSA1 was reassessed to Nifs (slow growth by nitrogen fixation) from our previous description of Nif- (Saeki, K., Suetsugu, Y., Tokuda, K., Miyatake, Y., Young, D. A., Marrs, B. L. and Matsubara, H. (1991) J. Biol. Chem. 266, 12889-12895). Substitution of Cys59 to Ser yielded an almost fully active product, while that of Cys54 did not. Gradual deletions and deletion-substitution of the 8 residues between Cys41 and Cys50 also yielded active products. Second, three of the modified FdxN proteins were subjected to purification. Only the GA protein, whose 8 residues between positions 42 and 49 were replaced by the Gly-Ala sequence, was purified. The GA protein and the authentic FdxN showed similar optical properties. The two clusters in the former had Em values of -490 and -430 mV, while those in the latter had an identical value ...Continue Reading

References

Jun 1, 1977·Journal of Biochemistry·T HaseM C Evans
Nov 7, 1975·Archives of Microbiology·P F WeaverH Gest
Jan 1, 1991·Advances in Protein Chemistry·J B Howard, D C Rees
Jan 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·A E MartínP J Stephens
Jan 1, 1987·Methods in Enzymology·J Vieira, J Messing
Jan 1, 1987·Methods in Enzymology·T A KunkelR A Zakour
Oct 24, 1995·Proceedings of the National Academy of Sciences of the United States of America·B ShenB K Burgess
Jul 28, 1995·Science·R D FleischmannJ M Merrick

❮ Previous
Next ❯

Citations

Oct 24, 1998·Progress in Biophysics and Molecular Biology·H Sticht, P Rösch
May 22, 2013·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Bradley Scott Perrin, Toshiko Ichiye
Nov 13, 2018·Protein Science : a Publication of the Protein Society·Stephanie J MaioccoSean J Elliott

❮ Previous
Next ❯

Related Concepts

Related Feeds

ASBMB Publications

The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.