Online immunocapture ICP-MS for the determination of the metalloprotein ceruloplasmin in human serum

Bernevic, Bogdan; El‐Khatib, Ahmed H.; Jakubowski, Norbert; Weller, Michael G.

doi:10.1186/s13104-018-3324-7

Cited by 21 publications

(23 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Using the example of ceruloplasmin, we checked the ability of the obtained clones to synthesize secretory cuproenzymes. Ceruloplasmin, a polyfunctional blue (ferr)oxidase of hepatic origin, is the main copper-containing blood serum protein [20][21][22]. It has been shown that ceruloplasmin is also synthesized in the lungs [23], its function has not been established.…”

Section: Resultsmentioning

confidence: 99%

CRISP-R/Cas9 Mediated Deletion of Copper Transport Genes CTR1 and DMT1 in NSCLC Cell Line H1299. Biological and Pharmacological Consequences

Ilyechova

Bonaldi

Orlov

et al. 2018

Preprint

View full text Add to dashboard Cite

Copper, the highly toxicity micronutrient, plays two essential roles: it is a catalytic and structural cofactor for Cu-dependent enzymes, and it acts as a secondary messenger. In the cells, copper is imported by CTR1, a transmembrane high-affinity copper importer, and DMT1 (divalent metal transporter). In cytosol, enzyme-specific chaperones receive copper from CTR1 C-terminus and deliver it to their apoenzymes. DMT1 cannot be a donor of catalytic copper because it does not have cytosol domain which is required for copper transfer to the Cu-chaperons and following to cuproenzymes. Here we assume that DMT1 can mediate copper way required for regulatory copper pool. To verify this thought, we used CRISPR/Cas9 to generate H1299 cell line with CTR1 or DMT1 single knockout (KO) and CTR1/DMT1 double knockout (DKO). To confirm KOs of the genes qRT-PCR were used. Two independent clones for each gene were selected for further studies. In CTR1-KO cells, expression of the DMT1 gene was significantly increased. In subcellular compartments, copper concentration decreased dramatically in DKO cells. CTR1-KO cells, but not DMT1-KO, demonstrated reduced sensitivity to cisplatin and silver ions, agents that enter the cell through CTR1. The expression of genes, whose protein products require copper: HIF1α, XIAP, COMMD1, CCS, Cp, but not SOD1 and NF-kB, changed their level. Perhaps these data will help to understand how the disturbances of copper homeodynamics lead to the development of neurodegenerative and oncological disorders. Possibility of using CTR1 KO and DMT1 KO cells to study homeodynamics of catalytic and signaling copper selectively is discussed.

show abstract

Section: Resultsmentioning

confidence: 99%

CRISP-R/Cas9 Mediated Deletion of Copper Transport Genes CTR1 and DMT1 in NSCLC Cell Line H1299. Biological and Pharmacological Consequences

Ilyechova

Bonaldi

Orlov

et al. 2018

Preprint

View full text Add to dashboard Cite

show abstract

“…For the isotopically labelled internal standard, a determination of the isotopic purity would be helpful. Thus, the efforts towards hepcidin-25 standardization [38] could be complemented by the development of a reference material additionally characterized by qNMR, ICP-MS [61] and amino acid analysis [53]. We would like to raise the awareness that synthetic and natural peptides can contain a large number of different proteoforms, from which isomers and metal complexes are particularly difficult to identify.…”

Section: Discussionmentioning

confidence: 99%

Investigations of the Copper Peptide Hepcidin-25 by LC-MS/MS and NMR

Abbas

Vranić

Hoffmann

et al. 2018

Preprint

Self Cite

View full text Add to dashboard Cite

Abstract:Hepcidin-25 was identified as the main iron regulator in the human body by binding to the sole ironexporter ferroportin. Studies showed that the N-terminus of hepcidin is responsible for this interaction, the same N-terminus that encompasses a small copper(II)-binding site known as ATCUN (amino terminal Cu(II)-and Ni(II)-binding) motif. Interestingly, this copper-binding property is largely ignored in most papers dealing with hepcidin-25. In this context, detailed investigations of the formed complex of hepcidin-25 with copper could reveal insights into its biological role. The present work is mainly focused on the study of the metal-bound form of hepcidin-25, which could be considered the biologically active form. The first part is devoted to the reversed-phase chromatographic separation of copper-bound and copper-free hepcidin-25, which was achieved by applying basic mobile phases containing 0.1% ammonia. Further, mass spectrometry (tandem mass spectrometry MS/MS, high resolution mass spectrometry HRMS) and nuclear magnetic resonance (NMR) spectroscopy were employed to characterize the copper-peptide. Lastly, a 3D model of hepcidin-25 with bound copper(II) is presented. The identification of metal complexes and potential isoforms and isomers, from which the latter usually are left undetected by mass spectrometry, led to the conclusion that complementary analytical methods are needed to characterize a peptide calibrant or reference material comprehensively. Quantitative nuclear magnetic resonance (qNMR), inductively-coupled plasma mass spectrometry (ICP-MS), ion-mobility spectrometry (IMS) and chiral amino acid analysis (AAA) should be considered among others.

show abstract

“…In the extracellular fluids of multicellular organisms, such as in the intracellular space, copper is coordinated by various carriers. Copper is found in ceruloplasmin (Cp), albumin, α-2-macroglobulin, and the bis-histidine complex [ 95 , 105 , 106 , 107 ]. In humans, more than 95% of copper is bound to Cp [ 105 ].…”

Section: Copper Turnover In the Body Of Adult Mammalsmentioning

confidence: 99%

“…Copper is found in ceruloplasmin (Cp), albumin, α-2-macroglobulin, and the bis-histidine complex [ 95 , 105 , 106 , 107 ]. In humans, more than 95% of copper is bound to Cp [ 105 ]. In mice, ceruloplasmin includes only about 60% of serum copper, and about 40% of copper is associated with α-2-macroglobulin [ 92 ].…”

Section: Copper Turnover In the Body Of Adult Mammalsmentioning

confidence: 99%

Copper Metabolism of Newborns Is Adapted to Milk Ceruloplasmin as a Nutritive Source of Copper: Overview of the Current Data

et al. 2018

View full text Add to dashboard Cite

Copper, which can potentially be a highly toxic agent, is an essential nutrient due to its role as a cofactor for cuproenzymes and its participation in signaling pathways. In mammals, the liver is a central organ that controls copper turnover throughout the body, including copper absorption, distribution, and excretion. In ontogenesis, there are two types of copper metabolism, embryonic and adult, which maintain the balance of copper in each of these periods of life, respectively. In the liver cells, these types of metabolism are characterized by the specific expression patterns and activity levels of the genes encoding ceruloplasmin, which is the main extracellular ferroxidase and copper transporter, and the proteins mediating ceruloplasmin metalation. In newborns, the molecular genetic mechanisms responsible for copper homeostasis and the ontogenetic switch from embryonic to adult copper metabolism are highly adapted to milk ceruloplasmin as a dietary source of copper. In the mammary gland cells, the level of ceruloplasmin gene expression and the alternative splicing of its pre-mRNA govern the amount of ceruloplasmin in the milk, and thus, the amount of copper absorbed by a newborn is controlled. In newborns, the absorption, distribution, and accumulation of copper are adapted to milk ceruloplasmin. If newborns are not breast-fed in the early stages of postnatal development, they do not have this natural control ensuring alimentary copper balance in the body. Although there is still much to be learned about the neonatal consequences of having an imbalance of copper in the mother/newborn system, the time to pay attention to this problem has arrived because the neonatal misbalance of copper may provoke the development of copper-related disorders.

show abstract

Online immunocapture ICP-MS for the determination of the metalloprotein ceruloplasmin in human serum

Cited by 21 publications

References 14 publications

CRISP-R/Cas9 Mediated Deletion of Copper Transport Genes CTR1 and DMT1 in NSCLC Cell Line H1299. Biological and Pharmacological Consequences

CRISP-R/Cas9 Mediated Deletion of Copper Transport Genes CTR1 and DMT1 in NSCLC Cell Line H1299. Biological and Pharmacological Consequences

Investigations of the Copper Peptide Hepcidin-25 by LC-MS/MS and NMR

Copper Metabolism of Newborns Is Adapted to Milk Ceruloplasmin as a Nutritive Source of Copper: Overview of the Current Data

Contact Info

Product

Resources

About