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Journal of Bacteriology, October 1999, p. 6524-6529, Vol. 181, No. 20
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Purification of P_II and P_II-UMP and In Vitro Studies of Regulation of Glutamine Synthetase in Rhodospirillum rubrum

Magnus Johansson and Stefan Nordlund^*

Department of Biochemistry, Arrhenius Laboratories for Natural Sciences, Stockholm University, S-106 91 Stockholm, Sweden

Received 9 March 1999/Accepted 27 July 1999

The P_II protein from Rhodospirillum rubrum was fused with a histidine tag, overexpressed in Escherichia coli, and purified by Ni²⁺-chelating chromatography. The uridylylated form of the P_II protein could be generated in E. coli. The effects on the regulation of glutamine synthetase by P_II, P_II-UMP, glutamine, and -ketoglutarate were studied in extracts from R. rubrum grown under different conditions. P_II and glutamine were shown to stimulate the ATP-dependent inactivation (adenylylation) of glutamine synthetase, which could be totally inhibited by -ketoglutarate. Deadenylylation (activation) of glutamine synthetase required phosphate, but none of the effectors studied had any major effect, which is different from their role in the E. coli system. In addition, deadenylylation was found to be much slower than adenylylation under the conditions investigated.

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^* Corresponding author. Mailing address: Department of Biochemistry, Arrhenius Laboratories for Natural Sciences, Stockholm University, S-106 91 Stockholm, Sweden. Phone: 46 8 16 29 32. Fax: 46 8 15 77 94. E-mail: stefan{at}biokemi.su.se.

Present address: Department of Biochemistry and Molecular Biology, James Cook University, Townsville, QLD 4811, Australia.

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Journal of Bacteriology, October 1999, p. 6524-6529, Vol. 181, No. 20
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

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