This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Rey, L Articles by Ruiz-Argüeso, T Search for Related Content PubMed PubMed Citation Articles by Rey, L Articles by Ruiz-Argüeso, T

 Previous Article  |  Next Article 

J Bacteriol. 1994 October; 176(19): 6066-6073

research-article

Purification of Rhizobium leguminosarum HypB, a nickel-binding protein required for hydrogenase synthesis.

Laboratorio de Microbiología, Escuela Técnica Superior de Ingenieros Agrónomos, Universidad Politécnica de Madrid, Spain.

ABSTRACT

The products of the Rhizobium leguminosarum hyp gene cluster are necessary for synthesis of a functional uptake [NiFe] hydrogenase system in symbiosis with pea plants, and at least for HypB and HypF, a role in hydrogenase-specific nickel metabolism has been postulated (L. Rey, J. Murillo, Y. Hernando, E. Hidalgo, E. Cabrera, J. Imperial, and T. Ruiz-Argüeso, Mol. Microbiol. 8:471-481, 1993). The R. leguminosarum hypB gene product has been overexpressed in Escherichia coli and purified by immobilized nickel chelate affinity chromatography in a single step. The purified recombinant HypB protein was able to bind 3.9 +/- 0.1 Ni2+ ions per HypB monomer in solution. Co2+, Cu2+, and Zn2+ ions competed with Ni2+ with increasing efficiency. Monospecific HypB antibodies were raised and used to show that HypB is synthesized in R. leguminosarum microaerobic vegetative cells and pea bacteroids but not in R. leguminosarum aerobic cells. HypB protein synthesized by R. leguminosarum microaerobic vegetative cells could also be isolated by immobilized nickel chelate affinity chromatography. A histidine-rich region at the amino terminus of the protein (23-HGHHHH DGHHDHDHDHDHHRGDHEHDDHHH-54) is proposed to play a role in nickel binding, both in solution and in chelated form.

This article has been cited by other articles:

• Leach, M. R., Zhang, J. W., Zamble, D. B. (2007). The Role of Complex Formation between the Escherichia coli Hydrogenase Accessory Factors HypB and SlyD. J. Biol. Chem. 282: 16177-16186 [Abstract] [Full Text]  
• Atanassova, A., Zamble, D. B. (2005). Escherichia coli HypA Is a Zinc Metalloprotein with a Weak Affinity for Nickel. J. Bacteriol. 187: 4689-4697 [Abstract] [Full Text]  
• Zhang, J. W., Butland, G., Greenblatt, J. F., Emili, A., Zamble, D. B. (2005). A Role for SlyD in the Escherichia coli Hydrogenase Biosynthetic Pathway. J. Biol. Chem. 280: 4360-4366 [Abstract] [Full Text]  
• Zambelli, B., Stola, M., Musiani, F., De Vriendt, K., Samyn, B., Devreese, B., Van Beeumen, J., Turano, P., Dikiy, A., Bryant, D. A., Ciurli, S. (2005). UreG, a Chaperone in the Urease Assembly Process, Is an Intrinsically Unstructured GTPase That Specifically Binds Zn2+. J. Biol. Chem. 280: 4684-4695 [Abstract] [Full Text]  
• Blokesch, M., Rohrmoser, M., Rode, S., Bock, A. (2004). HybF, a Zinc-Containing Protein Involved in NiFe Hydrogenase Maturation. J. Bacteriol. 186: 2603-2611 [Abstract] [Full Text]  
• Leech, H. K., Raux, E., McLean, K. J., Munro, A. W., Robinson, N. J., Borrelly, G. P. M., Malten, M., Jahn, D., Rigby, S. E. J., Heathcote, P., Warren, M. J. (2003). Characterization of the Cobaltochelatase CbiXL: EVIDENCE FOR A 4Fe-4S CENTER HOUSED WITHIN AN MXCXXC MOTIF. J. Biol. Chem. 278: 41900-41907 [Abstract] [Full Text]  
• Mehta, N., Olson, J. W., Maier, R. J. (2003). Characterization of Helicobacter pylori Nickel Metabolism Accessory Proteins Needed for Maturation of both Urease and Hydrogenase. J. Bacteriol. 185: 726-734 [Abstract] [Full Text]  
• Hube, M., Blokesch, M., Bock, A. (2002). Network of Hydrogenase Maturation in Escherichia coli: Role of Accessory Proteins HypA and HybF. J. Bacteriol. 184: 3879-3885 [Abstract] [Full Text]  
• Buhrke, T., Bleijlevens, B., Albracht, S. P. J., Friedrich, B. (2001). Involvement of hyp Gene Products in Maturation of the H2-Sensing [NiFe] Hydrogenase of Ralstonia eutropha. J. Bacteriol. 183: 7087-7093 [Abstract] [Full Text]  
• Jeon, W. B., Cheng, J., Ludden, P. W. (2001). Purification and Characterization of Membrane-associated CooC Protein and Its Functional Role in the Insertion of Nickel into Carbon Monoxide Dehydrogenase from Rhodospirillum rubrum. J. Biol. Chem. 276: 38602-38609 [Abstract] [Full Text]  
• Báscones, E., Imperial, J., Ruiz-Argüeso, T., Palacios, J. M. (2000). Generation of New Hydrogen-Recycling Rhizobiaceae Strains by Introduction of a Novel hup Minitransposon. Appl. Environ. Microbiol. 66: 4292-4299 [Abstract] [Full Text]  
• García-Domínguez, M., Lopez-Maury, L., Florencio, F. J., Reyes, J. C. (2000). A Gene Cluster Involved in Metal Homeostasis in the Cyanobacterium Synechocystis sp. Strain PCC 6803. J. Bacteriol. 182: 1507-1514 [Abstract] [Full Text]  
• Brito, B., Monza, J., Imperial, J., Ruiz-Argüeso, T., Palacios, J. M. (2000). Nickel Availability and hupSL Activation by Heterologous Regulators Limit Symbiotic Expression of the Rhizobium leguminosarum bv. Viciae Hydrogenase System in Hup- Rhizobia. Appl. Environ. Microbiol. 66: 937-942 [Abstract] [Full Text]  
• Watt, R. K., Ludden, P. W. (1998). The Identification, Purification, and Characterization of CooJ. A NICKEL-BINDING PROTEIN THAT IS CO-REGULATED WITH THE Ni-CONTAINING CO DEHYDROGENASE FROM RHODOSPIRILLUM RUBRUM. J. Biol. Chem. 273: 10019-10025 [Abstract] [Full Text]  
• Hottenrott, S., Schumann, T., Pluckthun, A., Fischer, G., Rahfeld, J.-U. (1997). The Escherichia coli SlyD Is a Metal Ion-regulated Peptidyl-prolyl cis/trans-Isomerase. J. Biol. Chem. 272: 15697-15701 [Abstract] [Full Text]  
• Brito, B., Martinez, M., Fernandez, D., Rey, L., Cabrera, E., Palacios, J. M., Imperial, J., Ruiz-Argueso, T. (1997). Hydrogenase genes from Rhizobium leguminosarum bv. viciae are controlled by the nitrogen fixation regulatory protein NifA. Proc. Natl. Acad. Sci. USA 94: 6019-6024 [Abstract] [Full Text]