Heavy metal-associated isoprenylated plant proteins (HIPPs) is an important metallochaperones due to its unique heavy metal binding domains (HMA) and the structural characteristics of isoprenylation motif. In order to identify the chelating ions of CsHIPP26.1 protein in Camellia sinensis (L.) cv. ‘Huangjinya’, the pET-32a-CsHIPP26.1 recombinant plasmids and empty carriers were respectively transferred into E. coli BL21, and then were cultured in LB liquid culture medium with 4 mol·L-1 single metal ions (CuCl2, ZnCl2, MgCl2, FeCl3, CaCl2) or multiple metal ions and 1 mmol·L-1 IPTG. The growth of E. coli in different ion media was observed, meanwhile the fusion target protein was obtained by His-tag protein purification magnetic bead. The contents of metal ions in fusion protein were analyzed by atomic absorption spectrophotometer, and the number of ions chelated by the protein was calculated. The results show that CsHIPP26.1 protein was only chelated with Zn2+ and Cu2+, and the chelating ability to Zn2+ was significantly higher than Cu2+. Based on the molar ratio of its bound metal ions to the target protein, the maximum number of Zn2+, Cu2+ chelated by CsHIPP26.1 protein was 2 and 1, respectively.
Key words
Camellia sinensis /
CsHIPP26.1 /
heavy metal-associated isoprenylated plant protein /
metal ions
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References
[1] 范延艮, 刘富浩, 赵秀秀, 等. 茶树’黄金芽’CsHIPP26.1基因克隆与光响应表达分析[J]. 植物生理学报, 2021, 57(5): 1087-1097.
Fan Y G, Liu F H, Zhao X X, et al.Cloning of CsHIPP26.1 in Camellia sinensis ‘Huangjinya’ and analysis of its expression level in response to light intensity[J]. Plant Physiology Journal, 2021, 57(5): 1087-1097.
[2] 范延艮. ‘黄金芽’茶树不同色泽新梢多组学比较及生理特性研究[D]. 泰安: 山东农业大学, 2019.
Fan Y G.Multiomics comparison and physiological characteristics of different colour shoots of Camellia sinensis var. Huangjingya [D]. Tai’an: Shandong Agricultural University, 2019.
[3] Neto J A, Turchetto-Zolet A C, Oliveira L, et al. Heavy metal-associated isoprenylated plant protein (HIPP): characterization of a family of proteins exclusive to plants[J]. The FEBS Journal, 2013, 280(7): 1604-1616.
[4] Dykema P E, Sipes P R, Marie A, et al.A new class of proteins capable of binding transition metals[J]. Plant Molecular Biology, 1999, 41(1): 139-150.
[5] Hung I H, Casareno R L B, Labesse G, et al. HAH1 is a copper-binding protein with distinct amino acid residues mediating copper homeostasis and antioxidant defense[J]. Journal of Biological Chemistry, 1998, 273(3): 1749-1754.
[6] Suzuki N, Yamaguchi Y, Koizumi N, et al.Functional characterization of a heavy metal binding protein CdI19 from Arabidopsis[J]. Plant Journal, 2010, 32(2): 165-173.
[7] Chandran D, Sharopova N, Ivashuta S, et al.Transcriptome profiling identified novel genes associated with aluminum toxicity, resistance and tolerance in Medicago truncatula[J]. Planta, 2008, 228(1): 151-166.
[8] Yamasaki H, Hayashi M, Fukazawa M, et al.SQUAMOSA promoter binding protein-like7 is a central regulator for copper homeostasis in Arabidopsis[J]. The Plant cell, 2018, 2009(1): 347-361.
[9] Jana K, Jan P, Vendula K, et al.Lack of phosphatidylglycerol inhibits chlorophyll biosynthesis at multiple sites and limits chlorophyllide reutilization in Synechocystis sp. Strain PCC 6803[J]. Plant Physiology, 2015, 169(2): 1307-1317.
[10] Gao H, Chen G, Han L, et al.Calcium influence on chilling resistance of grafting eggplant seedlings[J]. Journal of Plant Nutrition, 2005, 27(8): 1327-1339.
[11] 王孝平, 邢树礼. 考马斯亮蓝法测定蛋白含量的研究[J]. 天津化工, 2009, 23(3): 40-42.
Wang X P, Xing S L.Determination of protein quantitation using the method of coomassie brilliant blue[J]. Tianjin Chemical Industry, 2009, 23(3): 40-42.
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