Structure analysis of yeast glutaredoxin Grx6 protein produced in Escherichia coli

Abstract

BackgroundGrx6 is a yeast Golgi/endoplasmic reticulum protein involved in iron-sulfur binding that belongs to monothiol glutaredoxin-protein family. Grx6 has been biochemically characterized previously. Grx6 contains a conserved cysteine residue (Cys-136). Depending on the active-site sequences, Grxs can be classified to classic dithiol Grxs with a CXXC motif known as classes II and monothiol Grxs with a CXXS motif known as classes I, and Grx6 belongs to the class I with a CSYS motif.ResultsOur results showed how the loop between the N-terminal and C-terminal can affect the stability. When Grx6 was incubated with FeSO4·7H2O and (NH4)2Fe(SO4)2·6H2O, a disulfide bond was formed between the cysteine 136 and glutathione, and the concentration of dimer and tetramer was increased. The results presented various levels of stability of Grx6 with mutant and deleted amino acids. We also highlighted the difference between the monomer and dimer forms of the Grx6, in addition to comparison of the Fe-S cluster positions among holo forms of poplar Grx-C1, human Grx2 and Saccharomyces cerevisiae Grx6.ConclusionsIn this paper, we used a combination of spectroscopic and proteomic techniques to analyse the sequence and to determine the affected mutations and deletions in the stability of Grx6. Our results have increased the knowledge about the differences between monomer and dimer structures in cellular processes and proteins whose roles and functions depend on YCA1 in yeast.Electronic supplementary materialThe online version of this article (10.1186/s41021-018-0103-6) contains supplementary material, which is available to authorized users.

Divergence of active site motifs among different classes of Populus glutaredoxins results in substrate switches

“…Previous studies showed that some GRX proteins, such as Arabidopsis GRXC1, could form homodimers by ligating iron–sulfur clusters, leading to GRX inactivation (Riondet et al., 2012). When this kind of GRXs was expressed in E. coli , part of the recombinant GRXs existed as active monomeric proteins, and the other part as inactive iron–sulfur homodimers or/and tetramers (Abdalla et al., 2018; Lillig et al., 2005; Riondet et al., 2012). In this study, to obtain monomeric GRXs, all GRX proteins purified by an affinity chromatography column (Amylose Resin column or Nickel–Sepharose High Performance column) were further separated by size‐exclusion chromatography (Figure S3).…”

(Retracted) Image processing technology in Shanghai Stock Exchange options based on state-space model

“…It found that the definition of RBV is more extensive, and the study found that the estimator is more robust and more effective than the RV. Through empirical research on the data of the Shenzhen Stock Exchange and the Shanghai Composite Index, they confirmed that RBV has indeed achieved a more accurate estimation of stock market yield fluctuations 8 . By calculating the extremely poor price of high-frequency data, Christensen and Podolskij proposed another volatility estimator, the RRV estimator.…”

“…Through empirical research on the data of the Shenzhen Stock Exchange and the Shanghai Composite Index, they confirmed that RBV has indeed achieved a more accurate estimation of stock market yield fluctuations. 8 By calculating the extremely poor price of high-frequency data, Christensen and Podolskij proposed another volatility estimator, the RRV estimator. RRV has been used to reveal more useful information by exploiting the extremes of the entire price process.…”

A comprehensive mechanistic model of iron metabolism inSaccharomyces cerevisiae

“…193 It has glutathione-S-transferase and glutathione-dependent oxidoreductase activities. [215][216][217][218][219] Grx6 regulates the glutathionylation of proteins in the ER and Golgi, and controls the ratio of oxidized and reduced glutathione to protect the cell against oxidative stress.…”