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Journal of Bacteriology, May 1999, p. 2745-2751, Vol. 181, No. 9
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Role of XDHC in Molybdenum Cofactor Insertion into
Xanthine Dehydrogenase of Rhodobacter capsulatus
Silke
Leimkühler
and
Werner
Klipp*
Lehrstuhl für Biologie der
Mikroorganismen, Fakultät für Biologie,
Ruhr-Universität Bochum, D-44780 Bochum, Germany
Received 28 December 1998/Accepted 17 February 1999
Rhodobacter capsulatus xanthine dehydrogenase (XDH) is
composed of two subunits, XDHA and XDHB. Immediately downstream of xdhB, a third gene was identified, designated
xdhC, which is cotranscribed with xdhAB.
Interposon mutagenesis revealed that the xdhC gene product
is required for XDH activity. However, XDHC is not a subunit of active
XDH, which forms an
2
2 heterotetramer in
R. capsulatus. It was shown that XDHC neither is a
transcriptional regulator for xdh gene expression nor
influences XDH stability. To analyze the function of XDHC for XDH in
R. capsulatus, inactive XDH was purified from an
xdhC mutant strain. Analysis of the molybdenum cofactor
content of this enzyme demonstrated that in the absence of XDHC, no
molybdopterin cofactor MPT is present in the XDHAB tetramer. In
contrast, absorption spectra of inactive XDH isolated from the
xdhC mutant revealed the presence of iron-sulfur clusters and flavin adenine dinucleotide, demonstrating that XDHC is not required for the insertion of these cofactors. The absence of MPT from
XDH isolated from an xdhC mutant indicates that XDHC either
acts as a specific MPT insertase or might be a specific chaperone
facilitating the insertion of MPT and/or folding of XDH during or after
cofactor insertion.
*
Corresponding author. Mailing address: Lehrstuhl
für Biologie der Mikroorganismen, Fakultät für
Biologie, Ruhr-Universität Bochum, D-44780 Bochum, Germany.
Phone: 49-234-700-3100. Fax: 49-234-7094-620. E-mail:
werner.klipp{at}ruhr-uni-bochum.de.

Present address: Department of Biochemistry, Duke University
Medical Center, Durham, NC
27710.
Journal of Bacteriology, May 1999, p. 2745-2751, Vol. 181, No. 9
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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