Plant and Cell Physiology, 2003, Vol. 44, No. 10 1002-1012
© 2003 Oxford University Press
Novel Glyoxysomal Protein Kinase, GPK1, Identified by Proteomic Analysis of Glyoxysomes in Etiolated Cotyledons of Arabidopsis thaliana
1 Department of Cell Biology, National Institute for Basic Biology, Okazaki, 444-8585 Japan
2 Department of Molecular Biomechanics, School of Life Sciences, Graduate University for Advanced Studies, Okazaki, 444-8585 Japan
3 Department of Botany, Graduate School of Science, Kyoto University, Sakyo-ku, Kyoto, 606-8502 Japan
Glyoxysomes are present in etiolated cotyledons and contain enzymes for gluconeogenesis, which constitutes the major function of glyoxysomes. However, 281 genes seemingly related to peroxisomal functions occur in the Arabidopsis genome, implying that many unidentified proteins are present in glyoxysomes. To better understand the functions of glyoxysomes, we performed glyoxysomal proteomic analysis of etiolated Arabidopsis cotyledons. Nineteen proteins were identified as glyoxysomal proteins, including 13 novel proteins, one of which is glyoxysomal protein kinase 1 (GPK1). We cloned GPK1 cDNA by RT-PCR and characterized GPK1. The amino acid sequence deduced from GPK1 cDNA has a hydrophobic region, a putative protein kinase domain, and a possible PTS1 motif. Immunoblot analysis using fractions collected on a Percoll density gradient confirmed that GPK1 is localized in glyoxysomes. Analysis of suborganellar localization and protease sensitivity showed that GPK1 is localized on glyoxysomal membranes as a peripheral membrane protein and that the putative kinase domain is located inside the glyoxysomes. Glyoxysomal proteins are phosphorylated well in the presence of various metal ions and [g-32P]ATP, and one of them is identified as thiolase by immunoprecipitation. Immuno-inhibition of phosphorylation in glyoxysomes suggested that GPK1 phosphorylates a 40-kDa protein. These results show that protein phosphorylation systems are operating in glyoxysomes.
4 Present address: Department of Botany, Graduate School of Science, Kyoto University, Sakyo-ku, Kyoto, 606-8502 Japan.
5 Corresponding author: E-mail, mikosome{at}nibb.ac.jp; Fax, +81-564-55-7505.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  S. Reumann, S. Quan, K. Aung, P. Yang, K. Manandhar-Shrestha, D. Holbrook, N. Linka, R. Switzenberg, C. G. Wilkerson, A. P.M. Weber, et al. In-Depth Proteome Analysis of Arabidopsis Leaf Peroxisomes Combined with in Vivo Subcellular Targeting Verification Indicates Novel Metabolic and Regulatory Functions of Peroxisomes Plant Physiology, May 1, 2009; 150(1): 125 - 143. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. Arai, M. Hayashi, and M. Nishimura Proteomic Identification and Characterization of a Novel Peroxisomal Adenine Nucleotide Transporter Supplying ATP for Fatty Acid {beta}-Oxidation in Soybean and Arabidopsis PLANT CELL, December 1, 2008; 20(12): 3227 - 3240. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Eubel, E. H. Meyer, N. L. Taylor, J. D. Bussell, N. O'Toole, J. L. Heazlewood, I. Castleden, I. D. Small, S. M. Smith, and A. H. Millar Novel Proteins, Putative Membrane Transporters, and an Integrated Metabolic Network Are Revealed by Quantitative Proteomic Analysis of Arabidopsis Cell Culture Peroxisomes Plant Physiology, December 1, 2008; 148(4): 1809 - 1829. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. Ma and S. Reumann Improved prediction of peroxisomal PTS1 proteins from genome sequences based on experimental subcellular targeting analyses as exemplified for protein kinases from Arabidopsis J. Exp. Bot., October 1, 2008; 59(13): 3767 - 3779. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. Arai, M. Hayashi, and M. Nishimura Proteomic Analysis of Highly Purified Peroxisomes from Etiolated Soybean Cotyledons Plant Cell Physiol., April 1, 2008; 49(4): 526 - 539. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Reumann, L. Babujee, C. Ma, S. Wienkoop, T. Siemsen, G. E. Antonicelli, N. Rasche, F. Luder, W. Weckwerth, and O. Jahn Proteome Analysis of Arabidopsis Leaf Peroxisomes Reveals Novel Targeting Peptides, Metabolic Pathways, and Defense Mechanisms PLANT CELL, October 1, 2007; 19(10): 3170 - 3193. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. E. Lunn Compartmentation in plant metabolism J. Exp. Bot., January 1, 2007; 58(1): 35 - 47. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Ma, M. Haslbeck, L. Babujee, O. Jahn, and S. Reumann Identification and Characterization of a Stress-Inducible and a Constitutive Small Heat-Shock Protein Targeted to the Matrix of Plant Peroxisomes Plant Physiology, May 1, 2006; 141(1): 47 - 60. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. L. Heazlewood, J. Tonti-Filippini, R. E. Verboom, and A. H. Millar Combining Experimental and Predicted Datasets for Determination of the Subcellular Location of Proteins in Arabidopsis Plant Physiology, October 1, 2005; 139(2): 598 - 609. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. E. Turner, K. Greville, E. C. Murphy, and M. A. Hooks Characterization of Arabidopsis Fluoroacetate-resistant Mutants Reveals the Principal Mechanism of Acetate Activation for Entry into the Glyoxylate Cycle J. Biol. Chem., January 28, 2005; 280(4): 2780 - 2787. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Reumann, C. Ma, S. Lemke, and L. Babujee AraPerox. A Database of Putative Arabidopsis Proteins from Plant Peroxisomes Plant Physiology, September 1, 2004; 136(1): 2587 - 2608. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Reumann Specification of the Peroxisome Targeting Signals Type 1 and Type 2 of Plant Peroxisomes by Bioinformatics Analyses Plant Physiology, June 1, 2004; 135(2): 783 - 800. [Abstract] [Full Text] [PDF]
|
|