TY - JOUR
T1 - The role of tryptophan in the ferredoxin-dependent nitrite reductase of spinach
AU - Tripathy, Jatindra N.
AU - Hirasawa, Masakazu
AU - Kim, Sung Kun
AU - Setterdahl, Aaron T.
AU - Allen, James P.
AU - Knaff, David B.
N1 - Funding Information:
Acknowledgments The authors would like to thank Prof. Steven Rothstein (University of Guelph) for a generous gift of a spinach nitrite reductase clone, Prof. Toshiharu Hase (University of Osaka) for a generous gift of a clone of the E. coli cysG gene and for many helpful discussions. This research was supported by a grant (to DBK) from the U.S. Department of Energy (DE-FG02-99ER20346) and a grant (to JFA) from the U.S. Department of Agriculture (2003-02149).
PY - 2007/10
Y1 - 2007/10
N2 - A system has been developed for expressing a His-tagged form of the ferredoxin-dependent nitrite reductase of spinach in Escherichia coli. The catalytic and spectral properties of the His-tagged, recombinant enzyme are similar, but not identical, to those previously observed for nitrite reductase isolated directly from spinach leaf. A detailed comparison of the spectral, catalytic and fluorescence properties of nitrite reductase variants, in which each of the enzyme's eight tryptophan residues has been replaced using site-directed mutagenesis by either aromatic or non-aromatic amino acids, has been used to examine possible roles for tryptophan residues in the reduction of nitrite to ammonia catalyzed by the enzyme.
AB - A system has been developed for expressing a His-tagged form of the ferredoxin-dependent nitrite reductase of spinach in Escherichia coli. The catalytic and spectral properties of the His-tagged, recombinant enzyme are similar, but not identical, to those previously observed for nitrite reductase isolated directly from spinach leaf. A detailed comparison of the spectral, catalytic and fluorescence properties of nitrite reductase variants, in which each of the enzyme's eight tryptophan residues has been replaced using site-directed mutagenesis by either aromatic or non-aromatic amino acids, has been used to examine possible roles for tryptophan residues in the reduction of nitrite to ammonia catalyzed by the enzyme.
KW - Ferredoxin
KW - Nitrite reductase
KW - Tryptophan
UR - http://www.scopus.com/inward/record.url?scp=34548154052&partnerID=8YFLogxK
U2 - 10.1007/s11120-007-9198-5
DO - 10.1007/s11120-007-9198-5
M3 - Article
C2 - 17611813
AN - SCOPUS:34548154052
VL - 94
SP - 1
EP - 12
JO - Photosynthesis Research
JF - Photosynthesis Research
SN - 0166-8595
IS - 1
ER -