Abstract
Nonspecific lipid transfer protein (nsLTP) is usually basic and secreted low-molecular-mass protein in plants. The 3-D structure of nsLTP1 resembles that of elicitin produced by the plant pathogen Phytophthora cryptogea, which can bind to the plant plasma membrane putative receptor and activate the downstream responses. It is inferred that nsLTP1 may have similar binding sites on the plasma membranes. In this work, rice recombinant protein TRX-nsLTP110 labeled with 125I was shown to bind to rice plasma membrane preparations in a saturable curve, with an apparent Kd of 13.6 nM and Bmax of 150 fmol/mg proteins. Competition experiments revealed that the binding of TRX-nsLTP110 was specific, in contrast to the nonspecific binding of the fusion tag thioredoxin. Protease treatment assay showed that the binding sites were proteinaceous. Our results suggest that the binding sites of nsLTPs on plasma membranes may be ubiquitous in the plant kingdom. They may be competed out from the binding sites under pathogen attack, supporting a role for nsLTP1 in host defense response to pathogens.
Similar content being viewed by others
Abbreviations
- NsLTPs:
-
Nonspecific lipid transfer proteins
- SAR:
-
Systemic acquired resistance
- TRX:
-
Thioredoxin
- PVP:
-
Polyvinylpyrrolidone
- PMSF:
-
Phenylmethysulfenyl fluoride
- IC50 :
-
Inhibitory concentration
- TB:
-
Total binding
- SB:
-
Specific binding
- NSB:
-
Nonspecific binding
References
Deutch CE, Winicov I (1995) Post-transcriptional regulation of a salt-inducible alfalfa gene encoding a putative chimeric proline-rich cell wall protein. Plant Mol Biol 27:411–418
Jung HW, Hwang BK (2000) Isolation, partial sequencing, and expression of pathogenesis-related cDNA genes from pepper leaves infected by Xanthomonas campestris pv. vesicatoria. Mol Plant Microbe Interact 13:136–142
Gomes E, Sagot E, Gaillard C, Laquitaine L, Poinssot B, Sanejouand YH, Delrot S, Coutos-Thevenot P (2003) Nonspecific lipid-transfer protein genes expression in grape (Vitis sp.) cells in response to fungal elicitor treatments. Mol Plant Microbe Interact 16:456–464
Gomar J, Petit MC, Sodano P, Sy D, Marion D, Kader JC, Vovelle F, Ptak M (1996) Solution structure and lipid binding of a nonspecific lipid transfer protein extracted from maize seeds. Protein Sci 5:565–577
Da Silva P, Landon C, Industri B, Marais A, Marion D, Ponchet M, Vovelle F (2005) Solution structure of a tobacco lipid transfer protein exhibiting new biophysical and biological features. Proteins 59:356–367
Lin KF, Liu YN, Hsu ST, Samuel D, Cheng CS, Bonvin AM, Lyu PC (2005) Characterization and structural analyses of nonspecific lipid transfer protein 1 from mung bean. Biochemistry 44:5703–5712
Fefeu S, Bouaziz S, Huet JC, Pernollet JC, Guittet E (1997) Three-dimensional solution structure of beta cryptogein, a beta elicitin secreted by a phytopathogenic fungus Phytophthora cryptogea. Protein Sci 6:2279–2284
Motose H, Sugiyama M, Fukuda H (2004) A proteoglycan mediates inductive interaction during plant vascular development. Nature 429:873–878
Park SY, Jauh GY, Mollet JC, Eckard KJ, Nothnagel EA, Walling LL, Lord EM (2000) A lipid transfer-like protein is necessary for lily pollen tube adhesion to an in vitro stylar matrix. Plant Cell 12:151–164
Jung HW, Kim KD, Hwang BK (2005) Identification of pathogen-responsive regions in the promoter of a pepper lipid transfer protein gene (CALTPI) and the enhanced resistance of the CALTPI transgenic Arabidopsis against pathogen and environmental stresses. Planta 221:361–373
Wu G, Robertson AJ, Liu X, Zheng P, Wilen RW, Nesbitt NT, Gusta LV (2004) A lipid transfer protein gene BG-14 is differentially regulated by abiotic stress, ABA, anisomycin, and sphingosine in bromegrass (Bromus inermis). J Plant Physiol 161:449–458
Maldonado AM, Doerner P, Dixon RA, Lamb CJ, Cameron RK (2002) A putative lipid transfer protein involved in systemic resistance signalling in Arabidopsis. Nature 419:399–403
Ge XC, Chen JC, Lin Y, Sun CR, Cao KM (2002) Expression, purification and function of rice nonspecific lipid transfer protein. Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai) 34:83–87
Ge X, Chen J, Sun C, Cao K (2003) Preliminary study on the structural basis of the antifungal activity of a rice lipid transfer protein. Protein Eng 16:387–390
Bourque S, Ponchet M, Binet MN, Ricci P, Pugin A, Lebrun-Garcia A (1998) Comparison of binding properties and early biological effects of elicitins in tobacco cells. Plant Physiol 118:1317–1326
Wendehenne D, Binet MN, Blein JP, Ricci P, Pugin A (1995) Evidence for specific, high-affinity binding sites for a proteinaceous elicitor in tobacco plasma membrane. FEBS Lett 374:203–207
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
Widell S, Lundborg T, Larsson C (1982) Plasma membrane from oats prepared by partition in an aqueous polymer two-phase system: on the use of light-induced cytochrome b reduction as a marker for the plasma membrane. Plant Physiol 70:1429–1435
Bourque S, Binet MN, Ponchet M, Pugin A, Lebrun-Garcia A (1999) Characterization of the cryptogein binding sites on plant plasma membrane. J Biol Chem 274:34699–34705
Buhot N, Douliez JP, Jacquemard A, Marion D, Tran V, Maume BF, Milat ML, Ponchet M, Mikes V, Kader JC, Blein JP (2001) A lipid transfer protein binds to a receptor involved in the control of plant defence responses. FEBS Lett 509:27–30
Buhot N, Gomès E, Milat ML, Ponchet M, Marion D, Lequeu J, Delrot S, Coutos-Thévenot P, Blein JP (2004) Modulation of the biological activity of a tobacco LTP1 by lipid complexation. Mol Biol Cell 15:5047–5052
Durrant WE, Dong X (2004) Systemic acquired resistance. Annu Rev Phytopathol 42:185–209
Wang X (2004) Lipid signaling. Curr Opin Plant Biol 7:329–336
Acknowledgements
We thank Dr. Xingdang Liu, Xueping Zhang and Yun Lu from the Department of Nuclear Medicine of Huashan Hospital for the generous use of laboratory space and equipment and for helpful advice in handling radioactivity. We also thank Xiaocan Liu for technical assistance and Laura Heraty for revising the English. This work was supported by a research grant (30370297 to X. Ge) from the National Natural Science Foundation of China, and by the National Science Fund for Fostering Talents in Basic Research (J0630643 to the Biology Base of Fundan University).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Wang, X., Wang, H., Li, Y. et al. A rice lipid transfer protein binds to plasma membrane proteinaceous sites. Mol Biol Rep 36, 745–750 (2009). https://doi.org/10.1007/s11033-008-9238-z
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11033-008-9238-z