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 Pieper-Furst, U. Articles by Steinbuchel, A. Search for Related Content PubMed PubMed Citation Articles by Pieper-Furst, U. Articles by Steinbuchel, A.

 Previous Article  |  Next Article 

J. Bacteriol., May 1995, 2513-2523, Vol 177, No. 9
Copyright © 1995, American Society for Microbiology

Identification of the region of a 14-kilodalton protein of Rhodococcus ruber that is responsible for the binding of this phasin to polyhydroxyalkanoic acid granules

U Pieper-Furst, MH Madkour, F Mayer and A Steinbuchel
Institut fur Mikrobiologie, Georg-August-Universitat Gottingen, Germany.

The function of the polyhydroxyalkanoic acid (PHA) granule-associated GA14 protein of Rhodococcus ruber was investigated in Escherichia coli XL1-Blue, which coexpressed this protein with the polyhydroxybutyric acid (PHB) biosynthesis operon of Alcaligenes eutrophus. The GA14 protein had no influence on the biosynthesis rate of PHB in E. coli XL1- Blue(pSKCO7), but this recombinant E. coli strain formed smaller PHB granules than were formed by an E. coli strain that expressed only the PHB operon. Immunoelectron microscopy with GA14-specific antibodies demonstrated the binding of GA14 protein to these mini granules. In a previous study, two hydrophobic domains close to the C terminus of the GA14 protein were analyzed, and a working hypothesis that suggested an anchoring of the GA14 protein in the phospholipid monolayer surrounding the PHA granule core by these hydrophobic domains was developed (U. Pieper-Furst, M. H. Madkour, F. Mayer, and A. Steinbuchel, J. Bacteriol. 176:4328-4337, 1994). This hypothesis was confirmed by the construction of C-terminally truncated variants of the GA14 protein lacking the second or both hydrophobic domains and by the demonstration of their inability to bind to PHB granules. Further confirmation of the hypothesis was obtained by the construction of a fusion protein composed of the acetaldehyde dehydrogenase II of A. eutrophus and the C terminus of the GA14 protein containing both hydrophobic domains and by its affinity to native and artificial PHB granules.

This article has been cited by other articles:

• Neumann, L., Spinozzi, F., Sinibaldi, R., Rustichelli, F., Potter, M., Steinbuchel, A. (2008). Binding of the Major Phasin, PhaP1, from Ralstonia eutropha H16 to Poly(3-Hydroxybutyrate) Granules. J. Bacteriol. 190: 2911-2919 [Abstract] [Full Text]  
• Dennis, D., Sein, V., Martinez, E., Augustine, B. (2008). PhaP Is Involved in the Formation of a Network on the Surface of Polyhydroxyalkanoate Inclusions in Cupriavidus necator H16. J. Bacteriol. 190: 555-563 [Abstract] [Full Text]  
• Yamada, M., Yamashita, K., Wakuda, A., Ichimura, K., Maehara, A., Maeda, M., Taguchi, S. (2007). Autoregulator Protein PhaR for Biosynthesis of Polyhydroxybutyrate [P(3HB)] Possibly Has Two Separate Domains That Bind to the Target DNA and P(3HB): Functional Mapping of Amino Acid Residues Responsible for DNA Binding. J. Bacteriol. 189: 1118-1127 [Abstract] [Full Text]  
• Hanisch, J., Waltermann, M., Robenek, H., Steinbuchel, A. (2006). The Ralstonia eutropha H16 phasin PhaP1 is targeted to intracellular triacylglycerol inclusions in Rhodococcus opacus PD630 and Mycobacterium smegmatis mc2155, and provides an anchor to target other proteins.. Microbiology 152: 3271-3280 [Abstract] [Full Text]  
• Barnard, G. C., McCool, J. D., Wood, D. W., Gerngross, T. U. (2005). Integrated Recombinant Protein Expression and Purification Platform Based on Ralstonia eutropha. Appl. Environ. Microbiol. 71: 5735-5742 [Abstract] [Full Text]  
• Waltermann, M., Steinbuchel, A. (2005). Neutral Lipid Bodies in Prokaryotes: Recent Insights into Structure, Formation, and Relationship to Eukaryotic Lipid Depots. J. Bacteriol. 187: 3607-3619 [Full Text]  
• Banki, M. R., Gerngross, T. U., Wood, D. W. (2005). Novel and economical purification of recombinant proteins: Intein-mediated protein purification using in vivo polyhydroxybutyrate (PHB) matrix association. Protein Sci. 14: 1387-1395 [Abstract] [Full Text]  
• Schultheiss, D., Handrick, R., Jendrossek, D., Hanzlik, M., Schuler, D. (2005). The Presumptive Magnetosome Protein Mms16 Is a Poly(3-Hydroxybutyrate) Granule-Bound Protein (Phasin) in Magnetospirillum gryphiswaldense. J. Bacteriol. 187: 2416-2425 [Abstract] [Full Text]  
• Potter, M., Muller, H., Steinbuchel, A. (2005). Influence of homologous phasins (PhaP) on PHA accumulation and regulation of their expression by the transcriptional repressor PhaR in Ralstonia eutropha H16. Microbiology 151: 825-833 [Abstract] [Full Text]  
• Moldes, C., Garcia, P., Garcia, J. L., Prieto, M. A. (2004). In Vivo Immobilization of Fusion Proteins on Bioplastics by the Novel Tag BioF. Appl. Environ. Microbiol. 70: 3205-3212 [Abstract] [Full Text]  
• Korotkova, N., Chistoserdova, L., Lidstrom, M. E. (2002). Poly-{beta}-Hydroxybutyrate Biosynthesis in the Facultative Methylotroph Methylobacterium extorquens AM1: Identification and Mutation of gap11, gap20, and phaR. J. Bacteriol. 184: 6174-6181 [Abstract] [Full Text]  
• Potter, M., Madkour, M. H., Mayer, F., Steinbuchel, A. (2002). Regulation of phasin expression and polyhydroxyalkanoate (PHA) granule formation in Ralstonia eutropha H16. Microbiology 148: 2413-2426 [Abstract] [Full Text]  
• Maehara, A., Taguchi, S., Nishiyama, T., Yamane, T., Doi, Y. (2002). A Repressor Protein, PhaR, Regulates Polyhydroxyalkanoate (PHA) Synthesis via Its Direct Interaction with PHA. J. Bacteriol. 184: 3992-4002 [Abstract] [Full Text]  
• York, G. M., Stubbe, J., Sinskey, A. J. (2002). The Ralstonia eutropha PhaR Protein Couples Synthesis of the PhaP Phasin to the Presence of Polyhydroxybutyrate in Cells and Promotes Polyhydroxybutyrate Production. J. Bacteriol. 184: 59-66 [Abstract] [Full Text]  
• York, G. M., Stubbe, J., Sinskey, A. J. (2001). New Insight into the Role of the PhaP Phasin of Ralstonia eutropha in Promoting Synthesis of Polyhydroxybutyrate. J. Bacteriol. 183: 2394-2397 [Abstract] [Full Text]  
• Klinke, S., de Roo, G., Witholt, B., Kessler, B. (2000). Role of phaD in Accumulation of Medium-Chain-Length Poly(3-Hydroxyalkanoates) in Pseudomonas oleovorans. Appl. Environ. Microbiol. 66: 3705-3710 [Abstract] [Full Text]  
• Maehara, A., Ueda, S., Nakano, H., Yamane, T. (1999). Analyses of a Polyhydroxyalkanoic Acid Granule-Associated 16-Kilodalton Protein and Its Putative Regulator in the pha Locus of Paracoccus denitrificans. J. Bacteriol. 181: 2914-2921 [Abstract] [Full Text]  
• Madison, L. L., Huisman, G. W. (1999). Metabolic Engineering of Poly(3-Hydroxyalkanoates): From DNA to Plastic. Microbiol. Mol. Biol. Rev. 63: 21-53 [Abstract] [Full Text]  
• Prieto, M. A., Bühler, B., Jung, K., Witholt, B., Kessler, B. (1999). PhaF, a Polyhydroxyalkanoate-Granule-Associated Protein of Pseudomonas oleovorans GPo1 Involved in the Regulatory Expression System for pha Genes. J. Bacteriol. 181: 858-868 [Abstract] [Full Text]  
• McCool, G. J., Cannon, M. C. (1999). Polyhydroxyalkanoate Inclusion Body-Associated Proteins and Coding Region in Bacillus megaterium. J. Bacteriol. 181: 585-592 [Abstract] [Full Text]