Crystal Structure of the Human Copper Chaperone ATOX1 Bound to Zinc Ion

Mangini, Vincenzo; Belviso, Benny Danilo; Nardella, Maria Incoronata; Natile, Giovanni; Arnesano, Fabio; Caliandro, Rocco

doi:10.3390/biom12101494

Cited by 6 publications

(2 citation statements)

References 53 publications

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“…In addition to their natural metal cofactor, purified recombinant proteins can bind to other metal ions that can maintain or even stabilize their structure. In this sense, Cu(I) has been replaced by Zn(II) in human and cyanobacterial copper chaperones for structural studies [ 66 , 67 , 68 ]. Thus, in addition to fixing the copper loading state of our purified ETRs, we also used zinc as a replacement in our LCP crystallization trials.…”

Section: Resultsmentioning

confidence: 99%

Crystallization of Ethylene Plant Hormone Receptor—Screening for Structure

Rüffer,

Thielmann,

Lemke

et al. 2024

Biomolecules

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The plant hormone ethylene is a key regulator of plant growth, development, and stress adaptation. Many ethylene-related responses, such as abscission, seed germination, or ripening, are of great importance to global agriculture. Ethylene perception and response are mediated by a family of integral membrane receptors (ETRs), which form dimers and higher-order oligomers in their functional state as determined by the binding of Cu(I), a cofactor to their transmembrane helices in the ER-Golgi endomembrane system. The molecular structure and signaling mechanism of the membrane-integral sensor domain are still unknown. In this article, we report on the crystallization of transmembrane (TM) and membrane-adjacent domains of plant ethylene receptors by Lipidic Cubic Phase (LCP) technology using vapor diffusion in meso crystallization. The TM domain of ethylene receptors ETR1 and ETR2, which is expressed in E. coli in high quantities and purity, was successfully crystallized using the LCP approach with different lipids, lipid mixtures, and additives. From our extensive screening of 9216 conditions, crystals were obtained from identical crystallization conditions for ETR1 (aa 1-316) and ETR2 (aa 1-186), diffracting at a medium–high resolution of 2–4 Å. However, data quality was poor and not sufficient for data processing or further structure determination due to rotational blur and high mosaicity. Metal ion loading and inhibitory peptides were explored to improve crystallization. The addition of Zn(II) increased the number of well-formed crystals, while the addition of ripening inhibitory peptide NIP improved crystal morphology. However, despite these improvements, further optimization of crystallization conditions is needed to obtain well-diffracting, highly-ordered crystals for high-resolution structural determination. Overcoming these challenges will represent a major breakthrough in structurally determining plant ethylene receptors and promote an understanding of the molecular mechanisms of ethylene signaling.

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Section: Resultsmentioning

confidence: 99%

Crystallization of Ethylene Plant Hormone Receptor—Screening for Structure

Rüffer,

Thielmann,

Lemke

et al. 2024

Biomolecules

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“…[ 34 ] ATOX1 captures cytosolic copper and subsequently transfers it to copper pumps in the trans-Golgi network, thereby facilitating copper availability to various copper-dependent oxidoreductases. [ 35 ] ATP7B is a copper transporter P1B-type ATPase that maintains intracellular copper ion homeostasis. [ 36 ] The expression of FDX1 was positively correlated with the level of infiltration of CD8+ T cells and neutrophils.…”

Section: Discussionmentioning

confidence: 99%

Identification of 6 cuproptosis-related genes for active ulcerative colitis with both diagnostic and therapeutic values

Zou,

Zhang,

Zhu

et al. 2023

Medicine

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Cuproptosis has been reported to affect a variety of diseases. Therefore, we aimed to examine the role of cuproptosis-related genes in active ulcerative colitis (UC). We acquired 2 datasets of active UC from the Gene Expression Omnibus database and created immune cell infiltrations to research immune cell dysregulation. Based on the cuproptosis gene set and differentially expressed genes (DEGs), we identified the differentially expressed genes of cuproptosis (CuDEGs). We then used 2 machine learning methods to screen hub CuDEGs. Subsequently, we performed validation on additional datasets and investigated the relationship between hub CuDEGs and drug treatments. Thirty-five controls with inactive UC and 90 patients with active UC were obtained from the training sets. A total of 9157 DEGs and 27 CuDEGs were identified, respectively. Immune cell infiltration analysis revealed that patients with active UC exhibited higher levels of activated dendritic cells and neutrophils as well as lower levels of CD8+ T cells, regulatory T cells (Tregs), and macrophage M2. A six-gene cuproptosis signature was identified using machine learning algorithms. We further validated that the 6 hub CuDEGs showed a strong correlation with active UC and acted as cuproptosis-related biomarkers of active UC. Moreover, the expression of ATOX1 was downregulated, and SUMF1, MT1G, ATP7B, FDX1, and LIAS expression was upregulated in the colonic mucosa of active UC patients who responded to golimumab or vedolizumab therapy. With the exception of ATP7B, the expression patterns of hub CuDEGs before and after infliximab treatment of patients with active UC were similar to those of golimumab and vedolizumab. Cuproptosis and active UC have a complex relationship, as illustrated in our study. ATOX1, SUMF1, MT1G, ATP7B, FDX1, and LIAS are cuproptosis-related hub genes of active UC. Our study opens new avenues for investigating UC progression and developing novel therapeutic potential targets for the disease.

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