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BcLTP, a novel lipid transfer protein in Brassica chinensis, may secrete and combine extracellular CaM

  • Physiology and Biochemistry
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Abstract

Lipid transfer proteins in plants are believed to be involved in many processes of cell physiology and development. In this work, a full-length cDNA encoding a novel lipid transfer protein, designated BcLTP was isolated from Brassica chinensis. At least two copies of BcLTP are present in whole genome of B. chinensis, and its transcripts preferably accumulate in second-year organs, implying its role in reproductive growth stage. The 118 amino acid sequence deduced from a 354 bp open reading frame (ORF) shares common features with other members of plants LTPs family. A putative signal peptide at the N terminus was tested for secretion function by the yeast signal sequence trap (YSST) system, and further confirmed by vesicular and extracellular localization of YFP fusion protein. A highly conserved CaM binding site at C terminus was found and the binding properties with two representative CaM isoforms, one is convergent AtCaM2, one is divergent SCaM5, were determined by gel overlay. We found that convergent AtCaM2 prefer high concentration of Ca2+ for binding BcLTP, while SCaM5 does not depend on Ca2+ concentration too much for binding BcLTP. The lipid binding feature of BcLTP was demonstrated using florescence-marked 1-pyrenedodecanoic acid, which can be enhanced by AtCaM2 in Ca2+ dependent manner and by SCaM5 in either presence or absence of Ca2+. The collected data suggest that BcLTP may secrete and combine extracellular CaM isoforms, which in turn, facilitate lipid binding of BcLTP via Ca2+ mediated signaling.

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Abbreviations

AtCaM2:

Arabidopsis calmodulin isoform 2

BAA:

Basic amphiphilic α-helix

BcLTP:

Brassica chinensis lipid transfer protein

ΔBcLTP :

Mature BcLTP coding sequence

CaM:

Calmodulin

DAB:

3,3′-Diaminobenzidine

IPTG:

Isoporopylthio-β-d-thiogalactoside

nWAK2 :

N-terminal WAK2

Pyr-12C:

1-Pyrenedodecanoic acid

SCaM5:

Soybean calmodulin isoform 5

YSST:

Yeast signal sequence trap

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Acknowledgments

The authors thank Dr. R.E Zielinski for providing plasmid ACaM2 cDNA, Prof. M. J. Cho for providing SCaM5 cDNA, Professor X. M. Chen for providing pAVA321 vector, Dr. K. A. Jacob for providing YSST system. This work was supported by Nation Science Foundation of China (No. 90208004 and 30370733), National Key Basic Research Special Fund of China grants (2006CB100700 and 2006CB910600).

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Authors and Affiliations

  1. Institute of Molecular Cell Biology, Hebei Normal University, Shijiazhuang, 050016, China

    Chunming Wang, Guohong Mao, Daye Sun & Ying Sun

  2. College of Life Science, Nankai University, Tianjin, 300071, China

    Wanqin Xie, Fang Chi, Wenquan Hu & Cuifeng Li

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  1. Chunming Wang
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  2. Wanqin Xie
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  3. Fang Chi
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  4. Wenquan Hu
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  5. Guohong Mao
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  6. Daye Sun
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  7. Cuifeng Li
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  8. Ying Sun
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Correspondence to Cuifeng Li or Ying Sun.

Additional information

Communicated by Y. Lu.

BcLTP (accession number in Genbank: EF216852).

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Wang, C., Xie, W., Chi, F. et al. BcLTP, a novel lipid transfer protein in Brassica chinensis, may secrete and combine extracellular CaM. Plant Cell Rep 27, 159–169 (2008). https://doi.org/10.1007/s00299-007-0434-4

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  • DOI: https://doi.org/10.1007/s00299-007-0434-4

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