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Originally published In Press as doi:10.1074/jbc.M204581200 on August 9, 2002

J. Biol. Chem., Vol. 277, Issue 42, 40106-40111, October 18, 2002
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Accumulation of 99Mo-containing Iron-Molybdenum Cofactor Precursors of Nitrogenase on NifNE, NifH, and NifX of Azotobacter vinelandii*

Priya RangarajDagger § and Paul W. LuddenDagger ||

From the  Department of Biochemistry and the Dagger  Center for the Study of Nitrogen Fixation, College of Agricultural and Life Sciences, University of Wisconsin-Madison, Madison, Wisconsin 53706

The biosynthesis of the iron-molybdenum cofactor (FeMo-co) of nitrogenase was investigated using the purified in vitro FeMo-co synthesis system and 99Mo. The purified system involves the addition of all components that are known to be required for FeMo-co synthesis in their purified forms. Here, we report the accumulation of a 99Mo-containing FeMo-co precursor on NifNE. Apart from NifNE, NifH and NifX also accumulate 99Mo label. We present evidence that suggests NifH may serve as the entry point for molybdenum incorporation into the FeMo-co biosynthetic pathway. We also present evidence suggesting a role for NifX in specifying the organic acid moiety of FeMo-co.

* This work has been supported by National Institutes of Health/NIGMS Grant GM35332 (to P. W. L.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

§ Present address: Pierce Chemical Co., 3747 N. Meridian Road, Rockford, IL 61105.

|| To whom correspondence should be addressed: College of Natural Resources, University of California, Berkeley, 101 Giannini Hall, Rm. 3100, Berkeley, CA 94720. Tel.: 510-642-7171; Fax: 510-642-4612; E-mail: pludden@nature.berkeley.edu.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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