doi:

DOI: 10.3724/SP.J.1249.2017.05457

Journal of Shenzhen University Science and Engineering (深圳大学学报理工版) 2017/34:5 PP.457-463

Characteristics of antioxidant activity of soybean PM1 protein and enhancement of tolerance of recombinant yeast to copper stress


Abstract:
Cu2+ is an essential micronutrient for plant growth, but it is toxic when plant growth under excess copper stress. Soybean GmPM1 protein belongs to late embryogenesis abundant (LEA) group 4 (LEA4) proteins, which has a high proportion of histidine residues in the protein sequence. Firstly, we investigate the protective function and mechanisms of GmPM1 protein in plant under Cu2+ stress. The leaves of soybean seedling are withered under 150 μmol/L CuSO4 stress, and at the meantime the expression of GmPM1 gene in the young leaves was up-regulated in 3 h and 24 h of the stress. Secondly, the yeast expression plasmid of pYES2-GmPM1 is constructed and then transformed into the copper-sensitive yeast mutant ΔYAP1 to create recombinants of ΔYAP1-GmPM1. The recombinant yeast expressing GmPM1 protein could enhance the tolerance to Cu2+ stress. Then, the activities of scavenging hydroxyl radicals of GmPM1 and GmPM1-C protein in vitro are determined by using Cu-ascorbic acid system, which is rich in histidine residual in their sequence. The results show that GmPM1 could chelate Cu2+ through histidine residual in the C-terminal of GmPM1 protein and exert the activity of scavenge hydroxyl radicals, thus could improve the tolerance of plants to Cu2+ stress.

Key words:soybean,GmPM1 protein,histidine,Cu2+ stress,scavenging hydroxyl radicals,recombinant yeast

ReleaseDate:2017-10-20 02:07:29



[1] 刘昀,刘国宝,李冉辉,等.胚胎晚期富集蛋白与生物的干旱胁迫耐受性[J]. 生物工程学报,2010, 26(5):569-575. Liu Yun, Liu Guobao, Li Ranhui, et al. Functions of late embryogenesis abundant proteins in desiccation-tolerance of organisms:a review[J]. Chinese Journal of Biotechnology, 2010, 26(5):569-575.(in Chinese)

[2] Garay-Arroyo A, Colmenero-Flores J M, Garciarrubio A, et al. Highly hydrophilic proteins in prokaryotes and eukaryotes are common during conditions of water deficit[J]. The Journal of Biological Chemistry, 2000, 275(8):5668-5674.

[3] Su Mengying, Huang Gan, Zhang Qing, et al. The LEA protein, ABR, is regulated by ABI5 and involved in dark-induced leaf senescence in Arabidopsis thaliana[J]. Plant Science, 2016, 247:93-103.

[4] Roberts J K, Desimone N a, Lingle W L, et al. Cellular concentrations and uniformity of cell-type accumulation of two lea proteins in cotton embryos[J]. The Plant Cell, 1993, 5(7):769-780.

[5] Olvera-Carrillo Y,Campos F,Reyes J L,et al.Functional analysis of the group 4 late embryogenesis abundant proteins reveals their relevance in the adaptive response during water deficit in Arabidopsis[J]. Plant Physiology, 2010, 154(1):373-390.

[6] Cuevas-Velazquez C L, Saab-Rincón G, Reyes J L, et al. The unstructured n-terminal region of Arabidopsis group 4 late embryogenesis abundant (LEA) proteins is required for folding and for chaperone-like activity under water deficit[J]. The Journal of Biological Chemistry, 2016, 291(20):10893-10903.

[7] Shih M D, Hsieh T Y, Lin T P, et al. Characterization of two soybean (Glycine max L.) LEA IV proteins by circular dichroism and Fourier transform infrared spectrometry[J]. Plant and Cell Physiology, 2010, 51(3):395-407.

[8] Liu Guobao, Liu Ke, Gao Yang, et al. Involvement of C-Terminal histidines in soybean PM1 protein oligomerization and Cu2+ binding[J]. Plant and Cell Physiology, 2017, 58(6):1018-1029.

[9] Wang Hui, Hu Tangjin, Huang Jianzi, et al. The expression of Millettia pinnata chalcone isomerase in Saccharomyces cerevisiae salt-sensitive mutants enhances salt-tolerance[J]. International Journal of Molecular Sciences, 2013, 14(5):8775-8786.

[10] Hara M, Kondo M, Kato T. A KS-type dehydrin and its related domains reduce Cu-promoted radical Generation and the histidine residues contribute to the radical-reducing activities[J]. Journal of Experimental Botany, 2013, 64(6):1615-1624.

[11] Park S H, Shalongo W, Stellwagen E. The role of PⅡ conformations in the calculation of peptide fractional helix content[J]. Protein Science, 1997, 6(8):1694-1700.

[12] Soulages J L, Kim K, Arrese E L, et al. Conformation of a group 2 late embryogenesis abundant protein from soybean. Evidence of poly(L-proline)-type Ⅱ structure[J]. Plant Physiology, 2003, 131(3):963-975.

[13] González-Mendoza D, Espadas Y Gil F, Escoboza-Garcia F, et al. Copper stress on photosynthesis of black mangle (Avicennia germinans)[J]. Anais da Academia Brasileira de Ciencias, 2013, 85(2):665-670.

[14] Chamseddine M, Wided B A, Guy H, et al. Cadmium and copper induction of oxidative stress and antioxidative response in tomato (Solanum lycopersicon) leaves[J]. Plant Growth Regulation, 2008, 57(1):89-99.

[15] 金枫,王翠,林海建,等.植物重金属转运蛋白研究进展[J]. 应用生态学报, 2010, 21(7):1875-1882. Jin Feng, Wang Cui, Lin Haijian, et al. Heavy metal-transport proteins in plants:a review[J]. Chinese Journal of Applied Ecology, 2010, 21(7):1875-1882.(in Chinese)

[16] Mu Peiqiang, Feng Dongru, Su Jianbin, et al. Cu2+ triggers reversible aggregation of a disordered His-rich dehydrin MpDhn12 from Musa paradisiacal[J]. Journal of Biochemistry, 2011, 150(5):491-499.

[17] Finkelstein R. Abscisic acid synthesis and response[J]. The Arabidopsis Book, 2013, 11(11):e0058.

[18] Mowla S B, Cuypers A, Driscoll S P, et al. Yeast complementation reveals a role for an Arabidopsis thaliana late embryogenesis abundant(LEA)-like protein in oxidative stress tolerance[J]. Plant Journal, 2006, 48(5):743-756.

[19] Tiffany M L, Krimm S. New chain conformations of poly(glutamic acid) and polylysine[J]. Biopolymers, 1968, 6(9):1379-1382.

[20] Rath A, Davidson A R, Deber C M. The structure of "unstructured" regions in peptides and proteins:role of the polyproline Ⅱ helix in protein folding and recognition[J]. Biopolymers, 2005, 80(2-3):179-185.

[21] Zou Yongdong, Hong Ruisha, He Shuwen, et al. Polyproline Ⅱ structure is critical for the enzyme protective function of soybean Em (LEA1) conserved domains[J]. Biotechnology Letters, 2011, 33(8):1667-1673.

[22] Rahman L N, Smith G S, Bamm V V, et al. Phosphorylation of Thellungiella salsuginea dehydrins TsDHN-1 and TsDHN-2 facilitates cation-induced conformational changes and actin assembly[J]. Biochemistry, 2011, 50(44):9587-9604.