Tang Y.,Shenzhen University |
Tang Y.,Shenzhen Key Laboratory of Microbial and Gene Engineering |
Ou Z.,Shenzhen University |
Ou Z.,The Key Laboratory for Marine Bioresource and Eco environmental Science |
And 4 more authors.
Biotechnology Letters | Year: 2014
Plant-specific BURP family proteins have a diverse subcellular localization with different functions. However, only limited studies have investigated the functions of their different domains. In the present study, the role of the N-terminal putative signal peptide in protein subcellular localization was investigated using a tobacco cell system. The results showed that SALI3-2 was present in vacuoles, whereas AtRD22 was directed to the apoplast. The N-terminal putative signal peptides of both proteins were confirmed to be the essential and critical domains for targeting the proteins to their destinations. We also demonstrate that the expression and accumulation of mGFP in tobacco cells was increased when mGFP was fused to the putative signal peptide of SALI3-2. The findings offer the potential application of this short peptide in protein production in plants. © 2014, Springer Science+Business Media Dordrecht. Source
Tang Y.-L.,Shenzhen Key Laboratory of Microbial and Gene Engineering |
Tang Y.-L.,Shenzhen University |
Gao Z.,Shenzhen Key Laboratory of Microbial and Gene Engineering |
Gao Z.,Shenzhen University |
And 8 more authors.
Gaodeng Xuexiao Huaxue Xuebao/Chemical Journal of Chinese Universities | Year: 2013
The effect of CuCl2 on the growth of the transgenic Arabidopsis carrying Sali3-2 gene was studied on 1/2 MS agar plates. The over-expression of Sali3-2 was elevated the tolerance of transgenic Arabidopsis seedling to Cu2+ stress. Sali3-2 gene was then cloned and the SALI3-2 protein deleted signal peptide was expressed and purified. SALI3-2 protein interaction with Cu2+ was further studied by immobilized metal ion affinity chromatography(IMAC). The results show that the protein can bind with Cu2+. The mechanism of Cu2+-SALI3-2 bonding was further analyzed by fuorescence spectrum and circular dichroism(CD) spectrum. Fluorescence analysis shows intrinsic fluorescence of SALI3-2 is quenched with the addition of Cu2+, which belongs to static fluorescence quenching. Relatively high affinity of SALI3-2 binding Cu2+ is revealed with a constant of 8.89×106 and a number of binding sites of 1.6. The secondary structure of SALI3-2 is not changed obviously with the addition of Cu2+ as shown in CD spectra. These results suggest that SALI3-2 is a Cu2+-binding protein. It may function by altering its conformation through binding Cu2+ to trigger the regulation mechanism and elevate the tolerance of cells to Cu2+ stress. The research provides the direct insight into molecular mechanism of the role of SALI3-2 in plant tolerance to Cu2+ stress. Source