Skip to main content
Log in

Cold stress changes the concanavalin A-positive glycosylation pattern of proteins expressed in the basal parts of rice leaf sheaths

  • Original Article
  • Published:
Amino Acids Aims and scope Submit manuscript

Abstract

Post-translational modifications such as glycosylation are important for changing the properties and functions of proteins. To analyze the importance of glycosylation during cold stress in rice, a proteomics approach was used. Proteins extracted from the basal part of rice leaf sheaths were separated by two-dimensional polyacrylamide gel electrophoresis, and subjected to lectin blot analysis using concanavalin A. From a total of 250 detected proteins, 22 reacted with the lectin, suggesting that they were N-glycosylated proteins. To determine how N-glycosylation of these proteins is affected by cold stress, rice seedlings were incubated at 5°C for 48 h, and proteins extracted from the basal parts of leaf sheaths were analyzed by the lectin blot assay. Cold stress changed the reactivity toward the lectin for 12 of the 22 glycoproteins. The identity of the 12 proteins was determined by protein sequencing and mass spectrometry with the majority of these glycoproteins being categorized as involved in energy production. Furthermore, calreticulin, one of the 12 glycoproteins, was also phosphorylated as a result of cold stress. These results indicate that cold stress of the basal parts of rice leaf sheaths changes the glycosylation and phosphorylation profiles of calreticulin, a key protein that regulates the quality control of other proteins.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Abbreviations

2,4-D:

2,4-Dichlorophenoxyacetic acid

2D-PAGE:

Two-dimensional polyacrylamide gel electrophoresis

IEF:

Isoelectric focusing

PVDF:

Polyvinylidene difluoride

CBB:

Coomassie brilliant blue

PBS:

Phosphate-buffered saline

MS:

Mass spectrometry

References

  • Abe T, Oka Y, Sasahara T (1994) Varietal variations in biochemical changes during growth and redifferentiation of rice callus cultures. Jpn J Genet 69:385–396

    Article  CAS  Google Scholar 

  • Bevan M, Bancroft I, Bent E, Love K et al (1998) Analysis of 1.9 Mb of contiguous sequence from chromosome 4 of Arabidopsis thaliana. Nature 391:485–488

    Article  PubMed  CAS  Google Scholar 

  • Chapman R, Sidrauski C, Walter P (1998) Intracellular signaling from the endoplasmic reticulum to the nucleus. Annu Rev Cell Dev Biol 14:459–485

    Article  PubMed  CAS  Google Scholar 

  • Cui S, Huang F, Wang J, Ma X, Cheng Y, Liu J (2005) A proteomic analysis of cold stress responses in rice seedlings. Proteomics 5:3162–3172

    Article  PubMed  CAS  Google Scholar 

  • Demirevska-Kepova K, Simova-Stoilova L, Kjurkchiev S (1999) Barley leaf rubisco binding protein and rubisco activase and their protein/protein interactions. Bulg J Plant Physiol 2:31–44

    Google Scholar 

  • Hashimoto M, Komatsu S (2007) Proteomic analysis of rice seedlings during cold stress. Proteomics 7:1293–1302

    Article  PubMed  CAS  Google Scholar 

  • Hayashi T, Yamada T, Nakayama K, Komatsu S, Koike S (2006) Susceptibity to coolness at the young microspore stage under high nitrogen supply in rice (Oryza sativa L.). Plant Prod Sci 9:212–218

    Article  CAS  Google Scholar 

  • Huang J, Zhang H, Wang J, Yang J (2003) Molecular cloning and characterization of rice 6-phosphogluconate dehydrogenase gene that is up-regulated by salt stress. Mol Biol Rep 30:223–227

    Article  PubMed  CAS  Google Scholar 

  • Iba K (2002) Acclimative response to temperature stress in higher plants: approaches of gene engineering for temperature tolerance. Annu Rev Plant Biol 53:225–245

    Article  PubMed  CAS  Google Scholar 

  • Imin N, Kerim T, Rolfe BG, Weinman JJ (2004) Effect of early cold stress on the maturation of rice anthers. Proteomics 4:1873–1882

    Article  PubMed  CAS  Google Scholar 

  • Kariana ME, Meyer DJ, Gibbon BC, Jung R, Boston RS (2005) Identification and characterization of endoplasmic reticulum-associated degradation proteins differentially affected by endoplasmic reticulum stress. Plant Physiol 138:218–231

    Article  Google Scholar 

  • Khan M, Takasaki H, Komatsu S (2005) Comprehensive phosphoproteome analysis in rice and identification of phosphoproteins responsive to different hormones/stresses. J Proteome Res 4:1592–1599

    Article  PubMed  CAS  Google Scholar 

  • Khush GS (1999) Green revolution: preparing for the 21st century. Genome 42:646–655

    Article  PubMed  CAS  Google Scholar 

  • Kim SH, Kim JY, Kim SJ, An KS, An G, Kim SR (2007) Isolation of cold stress-responsive genes in the reproductive organs, and characterization of the OsLti6b gene from rice (Oryza sativa L.). Plant Cell Rep (on line)

  • Kleczkowski LA, Geisler M, Ciereszko I, Johansson H (2004) UDP-glucose pytophosphorylase. An old protein with new tricks. Plant Physiol 134:912–918

    Article  PubMed  CAS  Google Scholar 

  • Koga-Ban Y, Abe M, Kitagawa Y (1991) Alteration in gene expression during cold treatment of rice plant. Plant Cell Physiol 31:901–905

    Google Scholar 

  • Komatsu S, Masuda T, Abe K, (1996) Phosphorylation of a protein (pp56) is related to the regeneration of rice cultured suspension cells. Plant Cell Physiol 37:748–753

    PubMed  CAS  Google Scholar 

  • Komatsu S, Konishi H, Hashimoto M (2007) The proteomics of plant cell membranes. J Exp Bot 58:103–112

    Article  PubMed  CAS  Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  PubMed  CAS  Google Scholar 

  • Li Z, Komatsu S (2000) Molecular cloning and characterization of calreticulin, a calcium-binding protein involved in the regeneration of rice cultured suspension cells. Eur J Biochem 267:737–745

    Article  PubMed  CAS  Google Scholar 

  • Li Z, Onodera H, Ugaki M, Tanaka H, Komatsu S (2003) Characterization of calreticulin as a phosphoprotein interacting with cold-induced protein kinase in rice. Biol Pharm Bull 26:256–261

    Article  PubMed  CAS  Google Scholar 

  • Lyons JM (1972) Phase transitions and control of cellular metabolism at low temperatures. Cryobiology 9:341–350

    Article  PubMed  CAS  Google Scholar 

  • Martinez I, Chrispeels MJ (2003) Genomic analysis of the unfolded protein response in Arabidopsis shows its connection to important cellular processes. Plant Cell 15:561–576

    Article  PubMed  CAS  Google Scholar 

  • Meng M, Geisler M, Johansson H, Mellerowicz EJ, Karpinski S, Kleczkowski LA (2007) Differential tissue/organ-dependent expression of two sucrose- and cold-responsive genes for UDP-glucose pyrophosphorylase in Populus. Gene 389:186–195

    Article  PubMed  CAS  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • O’Farrell PH (1975) High resolution two-dimensional electrophoresis of protein. J Biol Chem 250:4007–4021

    PubMed  CAS  Google Scholar 

  • Ohtsu I, Nakanishi T, Furuta M, Ando E, Nishimura O (2005) Direct matrix-assisted laser desorption/ionization time-of-flight mass spectrometric identification of proteins on membrane detected by Western blotting and lectin blotting. J Proteome Res 4:1391–1396

    Article  PubMed  CAS  Google Scholar 

  • Pedersen PL, Ko YH, Hong S (2000) ATP synthase in the year 2000: evolving views about the structures of these remarkable enzyme complexes. J Bioenerg Biomembr 32:325–332

    Article  PubMed  CAS  Google Scholar 

  • Rabbani MA, Maruyama K, Abe H, Khan MA, Katsura K, Ito Y, Yoshiwara K, Seki M, Shinozaki K, Yamaguchi-Shinozaki K (2003) Monitoring expression profiles of rice genes under cold, drought, and high-salinity stresses and abscisic acid application using cDNA microarray and RNA gel-blot analyses. Plant Physiol 133:1755–1767

    Article  PubMed  CAS  Google Scholar 

  • Raison JK, Lyons JM (1986) Chilling injury: a plea for uniform terminology. Plant Cell Environ 9:685–686

    Article  Google Scholar 

  • Rayon C, Lerouge P, Faye L (1998) The protein N-glycosylation in plants. J Exp Bot 49:1463–1472

    Article  CAS  Google Scholar 

  • de los Reyes BG, Morsy M, Gibbons J, Varma TS, Antoine W, McGrath JM, Halgren R, Redus M (2003) A snapshot of the low temperature stress transcriptome of developing rice seedlings (Oryza sativa L.) via ESTs from subtracted cDNA library. Theor Appl Genet 107:1071–1082

    Article  Google Scholar 

  • Saher S, Fernandez-Garcia N, Piqueras A, Hellin E, Olmos E (2005) Reducing properties, energy efficiency and carbohydrate metabolism in hyperhydric and normal carnation shoots cultured in vitro: a hypoxia stress? Plant Physiol Biochem 43:573–582

    Article  PubMed  CAS  Google Scholar 

  • Thomashow MF (1998) Role of cold-responsive genes in plant freezing tolerance. Plant Physiol 118:1–8

    Article  PubMed  CAS  Google Scholar 

  • Tyagi AK, Mohanty A (2000) Rice transformation for crop improvement and functional genomics. Plant Sci 158:1–18

    Article  PubMed  CAS  Google Scholar 

  • Urada R (2007) Cellular response to unfolded proteins in the endoplasmic reticulum of plants. FEBS J 274:1152–1171

    Article  Google Scholar 

  • Yan SP, Zhang QY, Tang ZC, Su WA, Sun WN (2006) Comparative proteomic analysis provides new insights into chilling stress responses in rice. Mol Cell Proteomics 5:484–496

    PubMed  CAS  Google Scholar 

  • Yu X, Peng YH, Zhang MH, Shao YJ, Su WA, Tang ZC (2006) Water relations and an expression analysis of plasma membrane intrinsic proteins in sensitive and tolerant rice during chilling and recovery. Cell Res 16:599–608

    Article  PubMed  Google Scholar 

  • Zhang X, Takano T, Liu S (2006) Identification of a mitochondrial ATP synthase small subunit gene (RMtATP6) expressed in response to salts and osmotic stresses in rice (Oryza sativa L.). J Exp Bot 57:193–200

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to S. Komatsu.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Komatsu, S., Yamada, E. & Furukawa, K. Cold stress changes the concanavalin A-positive glycosylation pattern of proteins expressed in the basal parts of rice leaf sheaths . Amino Acids 36, 115–123 (2009). https://doi.org/10.1007/s00726-008-0039-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00726-008-0039-4

Keywords

Navigation