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. 1970 Jan;65(1):137–144. doi: 10.1073/pnas.65.1.137

Formation of Assimilatory Nitrate Reductase by in vitro Inter-Cistronic Complementation in Neurospora crassa*

Alvin Nason 1, Alan D Antoine 1,†, Paul A Ketchum 1,‡, William A Frazier III 1, David K Lee 1
PMCID: PMC286202  PMID: 4391854

Abstract

In vitro complementation of the soluble assimilatory nicotinamide adenine dinucleotide phosphate, reduced form (NADPH)-nitrate reductase was attained by mixing cell-free preparations of certain Neurospora nitrate reductase mutants: induced nit-1 (uniquely possessing inducible NADPH-cytochrome c reductase) with (a) uninduced or induced nit-2 or nit-3, or (b) uninduced wild type. The complementing activity of induced nit-1 is soluble while that of nit-2, nit-3, and wild type is particulate but not of mitochondrial origin. All fractions are inactivated by heat or trypsin. The NADPH-nitrate reductase enzymes formed in the above three complementing mixtures are similar to the wild-type enzyme in sucrose density gradient profiles, molecular weight, substrate affinity, sensitivity to inhibitors and temperature, but show different ratios of associated enzyme activities. The data suggest that nitrate reductase consists of at least two protein subunits: a nitrate-inductible subunit as reflected by inductible NADPH-cytochrome c reductase, and a constitutive protein which is activated (as indicated by the appearance of flavine adenine dinucleotide, reduced form (FADH2)- and reduced methyl viologen-nitrate reductase activities) when it combines with the inductible subunit.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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