Chlorophyll Binding Ability of Non-chloroplastic DUF538 Protein Superfamily in Plants

  • Ashraf GholizadehEmail author
Research Article


The type-1 water soluble chlorophyll binding proteins (WSCP1) have been generally known as chlorophyll extractors and transporters from the thylakoid membrane to the chloroplast envelope, where the membrane bound chlorophyllase catabolizes the chlorophyll. Despite the type-2 WSCP, WSCP1 has been known to be located in the chloroplasts of the green plants. In the present study, the non-chloroplastic protein superfamily containing domain of unknown function 538 (DUF538) as functional homologue of type-1 WSCP has been identified in plants. The structural similarities/differences and the cellular locations of Celosia cristata DUF538 and Chenopodium album WSCP1 were predicted by using computational tools and the chlorophyll binding abilities of their purified maltose binding protein-fused forms were estimated by maltose binding affinity method. It was predicted that despite CaWSCP1, CcDUF538 is a non-chloroplastic protein. The chlorophyll binding abilities of the recombinant fusion forms of test WSCP1 and DUF538 were estimated to be about 58 and 56%, respectively. Considering DUF538 as stress-induced protein, it was speculated that they may form complex with chlorophyll molecules or their hydrolyzed products out of chloroplasts to proceed the chlorophyll breakdown and nitrogen/carbon recycling in stress-challenged plants.


Celosia Chenopodium Chlorophyll binding Water soluble 



Water soluble chlorophyll binding proteins


Domain of unknown function


Bactericidal permeability increasing




Nitro blue tetrazolium


5-bromo-1-chloro-3-indolyl phosphate


Dimethyl formamide


Proto chlorophyll


Ethylene diamine tetra acidic acid


Maltose-binding protein



The author of this paper is thankful to the Research Institute for Fundamental Sciences (RIFS), University of Tabriz for the financial support.

Compliance with Ethical Standards

Conflict of interest

There is no conflict of interest in this study.


  1. 1.
    Horigome D, Satoh H, Itoh N, Mitsunaga K, Oonishi I, Nakagawa A, Uchida A (2007) Structural mechanism and photoprotective function of water-soluble chlorophyll-binding protein. J Biol Chem 282:6525–6531CrossRefPubMedGoogle Scholar
  2. 2.
    Noguchi T, Kamimura Y, Inoue Y, Itoh S (1999) Photoconversion of a water-soluble chlorophyll protein from Chenopodium album: resonance raman and fourier transform infrared study of protein and pigment structures. Plant Cell Physiol 40:305–310CrossRefGoogle Scholar
  3. 3.
    Satoh H, Uchida A, Nakayama K, Okada M (2001) Water-soluble chlorophyll protein in Brassicaceae plants is a stress-induced chlorophyll-binding protein. Plant Cell Physiol 42:906–911CrossRefPubMedGoogle Scholar
  4. 4.
    Takahashi S, Yoshikawa M, Kamada A, Ohtsuki T, Uchida A, Nakayama K, Satoh H (2013) The photoconvertible water-soluble chlorophyll-binding protein of Chenopodium album is a member of DUF538, a superfamily that distributes in Embryophyta. J Plant Physiol 170:1549–1552CrossRefPubMedGoogle Scholar
  5. 5.
    Takahashi S, Aizawa K, Nakayama K, Satoh H (2015) Water soluble chlorophyll binding proteins from Arabidopsis thaliana and Raphanus sativus target the endoplasmic reticulum body. BMC Res Notes 8:365CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Takahashi S, Yanai H, Oka-Takayama Y, Zanma-Sohtome A, Fujiyama K, Uchida A, Nakayama K, Satoh H (2013) Molecular cloning, characterization and analysis of the intracellular localization of a water-soluble chlorophyll binding protein (WSCP) from Virginia pepperweed (Lepidium virginicum), a unique WSCP that preferentially binds chlorophyll b in vitro. Planta 238:1065–1080CrossRefPubMedGoogle Scholar
  7. 7.
    Yamada K, Hara-Nishimura I, Nishimura M (2011) Unique defense strategy by the endoplasmic reticulum body in plants. Plant Cell Physiol 52:2039–2049CrossRefPubMedGoogle Scholar
  8. 8.
    Yamada K, Nagano AJ, Nishina M, Hara-Nishimura I, Nishimura M (2013) Identification of two novel endoplasmic reticulum body-specific integral membrane proteins. Plant Physiol 161:108–120CrossRefPubMedGoogle Scholar
  9. 9.
    Gholizadeh A, BaghbanKohnehrouz B (2010) Identification of DUF538 cDNA clone from Celosia cristata expressed sequences of none stressed and stressed leaves. Russ J Plant Physiol 57:247–252CrossRefGoogle Scholar
  10. 10.
    Gholizadeh A (2011) Heterologous expression of stress-responsive DUF538 domain containing protein and its morpho-biochemical consequences. Protein J 30:351–358CrossRefPubMedGoogle Scholar
  11. 11.
    Gholizadeh A, Baghbankohnehrouz S (2013) DUF538 protein super family is predicted to be the potential homologue of bactericidal/permeability-increasing protein in plant system. Protein J 32:163–171CrossRefPubMedGoogle Scholar
  12. 12.
    Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685CrossRefPubMedGoogle Scholar
  13. 13.
    Moran R (1982) Formulae for determination of chlorophyllous pigments extracted with N, N-dimethylformamide. Plant Physiol 69:1376–1381CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Fang Z, Bouwkamp JC, Solomos T (1998) Chlorophyllase activities and chlorophyll degradation during leaf senescence in non-yellowing mutant and wild type of Phaseolus vulgaris L. J Exp Bot 49:503–510Google Scholar
  15. 15.
    Nakagami H, Sugiyama N, Mochida K, Daudi A, Yoshida Y, Toyoda T, Tomita M, Ishihama Y, Shirasu K (2010) Large-scale comparative phosphoproteomics identifies conserved phosphorylation sites in plants. Plant Physiol 153:1161–1174CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Annamalai P, Yanagihara S (1999) Identification and characterization of a heat-stress induced gene in cabbage encodes a kunitz type protease inhibitor. J Plant Physiol 155:226–233CrossRefGoogle Scholar
  17. 17.
    Downing WL, Mauxion F, Fauvarque MO, Reviron MP, de Vienne D, Vartanian N, Giraudat J (1992) A Brassica napus transcript encoding a protein related to the Künitz protease inhibitor family accumulates upon water stress in leaves, not in seeds. Plant J 2:685–693PubMedGoogle Scholar
  18. 18.
    Nishio N, Satoh H (1997) A water-soluble chlorophyll protein in cauliflower may be identical to BnD22, a drought-induced, 22-kilodalton protein in rapeseed. Plant Physiol 115:841–846CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Satoh H, Nakayama K, Okada M (1998) Molecular cloning and functional expression of a water-soluble chlorophyll protein, a putative carrier of chlorophyll molecules in cauliflower. J Biol Chem 273:30568–30575CrossRefPubMedGoogle Scholar
  20. 20.
    Takamiya KI, Tsuchiya T, Ohta H (2000) Degradation pathway (s) of chlorophyll: what has gene cloning revealed? Trends Plant Sci 5:426–431CrossRefPubMedGoogle Scholar
  21. 21.
    Tsuchiya T, Ohta H, Masuda T, Mikami B, Kita N, Shioi Y, Takamiya K (1997) Purification and characterization of two isozymes of chlorophyllase from mature leaves of Chenopodium album. Plant Cell Physiol 38:1026–1031CrossRefGoogle Scholar
  22. 22.
    Schenk N, Schelbert S, Kanwischer M, Goldschmidt EE, Dörmann P, Hörtensteiner S (2007) The chlorophyllases AtCLH1 and AtCLH2 are not essential for senescence-related chlorophyll breakdown in Arabidopsis thaliana. FEBS Lett 27:5517–5525CrossRefGoogle Scholar
  23. 23.
    Takahashi S, Yanai H, Nakamaru Y, Uchida A, Nakayama K, Satoh H (2012) Molecular cloning, characterization and analysis of the intracellular localization of a water-soluble chlorophyll-binding protein from Brussels sprouts (Brassica oleracea var. gemmifera). Plant Cell Physiol 53:879–891CrossRefPubMedGoogle Scholar

Copyright information

© The National Academy of Sciences, India 2016

Authors and Affiliations

  1. 1.Department of BiochemistryUniversity of TabrizTabrizIran

Personalised recommendations