茶树黄金芽CsHIPP26.1蛋白螯合离子的筛选与鉴定

doi:10.13305/j.cnki.jts.2022.02.006

Abstract

Abstract: 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: heavy metal-associated isoprenylated plant protein, CsHIPP26.1, metal ions, Camellia sinensis

CLC Number:

S571.1

LIU Fuhao, FAN Yangen, WANG Yu, MENG Fanyue, ZHANG Lixia. Screening and Identification of Chaperone CsHIPP26.1 Chelating Ions in Tea Cultivar ‘Huangjinya’[J]. Journal of Tea Science, 2022, 42(2): 179-186.

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Screening and Identification of Chaperone CsHIPP26.1 Chelating Ions in Tea Cultivar ‘Huangjinya’

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