Plant Physiol 99(2): 693-699
Expression in <em>Escherichia coli</em> of Cytochrome <em>c</em> Reductase Activity from a Maize NADH:Nitrate Reductase Complementary DNA <sup><a href="#fn1" rid="fn1" class=" fn">1</a></sup>
Abstract
A cDNA clone was isolated from a maize (Zea mays L. cv W64A×W183E) scutellum λgt11 library using maize leaf NADH:nitrate reductase Zmnr1 cDNA clone as a hybridization probe; it was designated Zmnr1S. Zmnr1S was shown to be an NADH:nitrate reductase clone by nucleotide sequencing and comparison of its deduced amino acid sequence to Zmnr1. Zmnr1S, which is 1.8 kilobases in length and contains the code for both the cytochrome b and flavin adenine dinucleotide domains of nitrate reductase, was cloned into the EcoRI site of the Escherichia coli expression vector pET5b and expressed. The cell lysate contained NADH:cytochrome c reductase activity, which is a characteristic partial activity of NADH:nitrate reductase dependent on the cytochrome b and flavin adenine dinucleotide domains. Recombinant cytochrome c reductase was purified by immunoaffinity chromatography on monoclonal antibody Zm2(69) Sepharose. The purified cytochrome c reductase, which had a major size of 43 kilodaltons, was inhibited by polyclonal antibodies for maize leaf NADH:nitrate reductase and bound these antibodies when blotted to nitrocellulose. Ultraviolet and visible spectra of oxidized and NADH-reduced recombinant cytochrome c reductase were nearly identical with those of maize leaf NADH:nitrate reductase. These two enzyme forms also had very similar kinetic properties with respect to NADH-dependent cytochrome c and ferricyanide reduction.
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- Barber MJ, Notton BA. Spinach Nitrate Reductase : Effects of Ionic Strength and pH on the Full and Partial Enzyme Activities. Plant Physiol. 1990 Jun;93(2):537–540.[PMC free article] [PubMed] [Google Scholar]
- Campbell WH, Kinghorn KR. Functional domains of assimilatory nitrate reductases and nitrite reductases. Trends Biochem Sci. 1990 Aug;15(8):315–319. [PubMed] [Google Scholar]
- Campbell WH, Remmler JL. Regulation of Corn Leaf Nitrate Reductase : I. Immunochemical Methods for Analysis of the Enzyme's Protein Component. Plant Physiol. 1986 Feb;80(2):435–441.[PMC free article] [PubMed] [Google Scholar]
- Cannons AC, Iida N, Solomonson LP. Expression of a cDNA clone encoding the haem-binding domain of Chlorella nitrate reductase. Biochem J. 1991 Aug 15;278(Pt 1):203–209.[PMC free article] [PubMed] [Google Scholar]
- Cheng CL, Dewdney J, Nam HG, den Boer BG, Goodman HM. A new locus (NIA 1) in Arabidopsis thaliana encoding nitrate reductase. EMBO J. 1988 Nov;7(11):3309–3314.[PMC free article] [PubMed] [Google Scholar]
- Choi HK, Kleinhofs A, An GH. Nucleotide sequence of rice nitrate reductase genes. Plant Mol Biol. 1989 Dec;13(6):731–733. [PubMed] [Google Scholar]
- Crawford NM, Campbell WH. Fertile Fields. Plant Cell. 1990 Sep;2(9):829–835.[PMC free article][Google Scholar]
- Gowri G, Campbell WH. cDNA Clones for Corn Leaf NADH:Nitrate Reductase and Chloroplast NAD(P):Glyceraldehyde-3-Phosphate Dehydrogenase : Characterization of the Clones and Analysis of the Expression of the Genes in Leaves as Influenced by Nitrate in the Light and Dark. Plant Physiol. 1989 Jul;90(3):792–798.[PMC free article] [PubMed] [Google Scholar]
- Gowri G, Kenis JD, Ingemarsson B, Redinbaugh MG, Campbell WH. Nitrate reductase transcript is expressed in the primary response of maize to environmental nitrate. Plant Mol Biol. 1992 Jan;18(1):55–64. [PubMed] [Google Scholar]
- Hyde GE, Campbell WH. High-level expression in Escherichia coli of the catalytically active flavin domain of corn leaf NADH:nitrate reductase and its comparison to human NADH:cytochrome B5 reductase. Biochem Biophys Res Commun. 1990 May 16;168(3):1285–1291. [PubMed] [Google Scholar]
- Hyde GE, Wilberding JA, Meyer AL, Campbell ER, Campbell WH. Monoclonal antibody-based immunoaffinity chromatography for purifying corn and squash NADH: nitrate reductases. Evidence for an interchain disulfide bond in nitrate reductase. Plant Mol Biol. 1989 Aug;13(2):233–246. [PubMed] [Google Scholar]
- Johnson JL, Bastian NR, Rajagopalan KV. Molybdopterin guanine dinucleotide: a modified form of molybdopterin identified in the molybdenum cofactor of dimethyl sulfoxide reductase from Rhodobacter sphaeroides forma specialis denitrificans. Proc Natl Acad Sci U S A. 1990 Apr;87(8):3190–3194.[PMC free article] [PubMed] [Google Scholar]
- Kay CJ, Barber MJ. Assimilatory nitrate reductase from Chlorella. Effect of ionic strength and pH on catalytic activity. J Biol Chem. 1986 Oct 25;261(30):14125–14129. [PubMed] [Google Scholar]
- Kubo Y, Ogura N, Nakagawa H. Limited proteolysis of the nitrate reductase from spinach leaves. J Biol Chem. 1988 Dec 25;263(36):19684–19689. [PubMed] [Google Scholar]
- Redinbaugh MG, Campbell WH. Adaptation of the dye-binding protein assay to microtiter plates. Anal Biochem. 1985 May 15;147(1):144–147. [PubMed] [Google Scholar]
- Redinbaugh MG, Campbell WH. Quaternary structure and composition of squash NADH:nitrate reductase. J Biol Chem. 1985 Mar 25;260(6):3380–3385. [PubMed] [Google Scholar]
- Schnorr KM, Juricek M, Huang CX, Culley D, Kleinhofs A. Analysis of barley nitrate reductase cDNA and genomic clones. Mol Gen Genet. 1991 Jul;227(3):411–416. [PubMed] [Google Scholar]
- Slice LW, Taylor SS. Expression of the catalytic subunit of cAMP-dependent protein kinase in Escherichia coli. J Biol Chem. 1989 Dec 15;264(35):20940–20946. [PubMed] [Google Scholar]
- Studier FW. Use of bacteriophage T7 lysozyme to improve an inducible T7 expression system. J Mol Biol. 1991 May 5;219(1):37–44. [PubMed] [Google Scholar]
- Studier FW, Rosenberg AH, Dunn JJ, Dubendorff JW. Use of T7 RNA polymerase to direct expression of cloned genes. Methods Enzymol. 1990;185:60–89. [PubMed] [Google Scholar]