Immobilized metal ion affinity electrophoresis

Goubran‐Botros, Hany; Nanak, Elizabeth; Nour, Jamal Abdul; Birkenmeir, Gerd; Vijayalakshmi, M.A.

doi:10.1016/0021-9673(92)80132-e

Cited by 16 publications

(8 citation statements)

References 18 publications

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“…As a typical example, a-2-HS-glycoprotein is a protein, which was completely retained by the IDA-Zn 21 column, and this retention behavior is in agreement with the findings reported in Ref. [20] in the sense that a-2-HS-glycoprotein has His residues that are favorable for its retention on IDA-Zn 21 . Also, it was reported that a-2-HSglycoprotein has similar dissociation constants for IDA-Zn 21 and IDA-Cu 21 columns [20].…”

Section: Highlights Of the Retention Of Some Typical Proteins On The supporting

confidence: 87%

“…[20] in the sense that a-2-HS-glycoprotein has His residues that are favorable for its retention on IDA-Zn 21 . Also, it was reported that a-2-HSglycoprotein has similar dissociation constants for IDA-Zn 21 and IDA-Cu 21 columns [20]. However, since it was placed first in the tandem series, the IDA-Zn 21 column captured all of the a-2-HS-glycoprotein, thus allowing more affinity sites on the IDA-Cu 21 column to be available for the retention of other proteins.…”

Section: Highlights Of the Retention Of Some Typical Proteins On The mentioning

confidence: 98%

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Reduction of protein concentration range difference followed by multicolumn fractionation prior to 2‐DE and LC‐MS/MS profiling of serum proteins

Selvaraju

Rassi

2011

Electrophoresis

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This article is concerned with the reduction of protein concentration range differences by the peptide beads library technology (ProteoMiner™ or "equalizer" technology), which in principle allows the enrichment of proteins to the same concentration level (i.e. protein equalizer) regardless of the original protein abundance in a given biological fluid such as human serum, which is the subject of our investigation. After the equalization step, the captured proteins from human serum were fractionated on a series of tandem monolithic columns with surface-bound iminodiacetic acid ligands to which three different metal ions, namely, Zn²+, Ni²+ and Cu²+ were immobilized to yield the so-called immobilized metal affinity chromatography columns. These three monolithic columns were connected to a reversed-phase column packed with polystyrene divinyl benzene beads. Aliquots taken from the four collected fractions from the four tandem columns were subsequently fractionated by 2-DE. Also, aliquots from the four collected fractions were tryptically digested and analyzed by LC-MS/MS. The strategy of subsequent fractionation on the four tandem columns after equalization allowed the identification of more proteins than simply using the equalization by ProteoMiner™ . The equalizer technology was compared to the immuno-subtraction approach. While the ProteoMiner™ technology is superior in terms of the overall number of captured proteins, it only complements the immuno-subtraction approach since the latter can capture the proteins that were not captured by the former.

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Section: Highlights Of the Retention Of Some Typical Proteins On The supporting

confidence: 87%

Section: Highlights Of the Retention Of Some Typical Proteins On The mentioning

confidence: 98%

Reduction of protein concentration range difference followed by multicolumn fractionation prior to 2‐DE and LC‐MS/MS profiling of serum proteins

Selvaraju

Rassi

2011

Electrophoresis

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“…Aqueous two-phase systems containing metal-IDA-poly(ethylene glycol) (PEG) in PEG-dextran and PEG-salt systems have been used not only for extracting proteins [50] but also for affinity partitioning of human blood cells such as erythrocytes [51] or lymphocytes [52]. Immobilized metal-ion affinity gel electrophoresis (IMAGE) on, e.g., PEG-IDA-Cu(II) in agarose gels [53], and immobilized metal-ion affinity capillary electrophoresis (IMACE) with soluble polymer-supported ligands [54] are examples of further applications of the same basic principle, although none of these techniques has become as popular as IMAC itself. Other application of metal-affinity-based protein adsorption is in biosensor technique.…”

Section: Applications Of Immobilized Metal-ion Affinity Beyond Imacmentioning

confidence: 99%

“…In the beginning, the ligand was immobilized onto agarose beads and incorporated in an agarose gel [60]. Later, ligands coupled to soluble polymers [53] and monomeric polymerizable derivatives of the ligands that could be copolymerized into polyacrylamide gel [61].…”

Section: Immobilized Metal-ion Affinity Gel Electrophoresismentioning

confidence: 99%

Immobilized metal-ion affinity systems for recovery and structure–function studies of proteins at molecular, supramolecular, and cellular levels

Prasanna

Vijayalakshmi

2010

Pure and Applied Chemistry

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Immobilized metal-ion affinity (IMA) adsorption is a collective term that is used to include all kinds of adsorptions where the metal ion serves as the characteristic and most essential part of adsorption center. Of all the IMA techniques, immobilized metal-affinity chromato graphy (IMAC) has been gaining popularity as the choice of purification technique for proteins. IMAC represents a separation technique that is primarily useful for proteins with natural surface exposed-histidine residues and for recombinant proteins with engineered histidine tag. This review also gives insight into other nonchromatographic applications of IMA adsorption such as immobilized metal-ion affinity gel electrophoresis (IMAGE), immobilized metal-ion affinity capillary electrophoresis (IMACE), and immobilized metal-ion affinity partitioning (IMAP).Keywords: immobilized metal-affinity chromato graphy; immobilized metal-ion affinity capillary electrophoresis; immobilized metal-ion affinity gel electrophoresis; immobilized metal-ion affinity partitioning; metal affinity; proteins.IMA adsorption is a collective term that is used to include all kinds of adsorptions where a metal ion, with affinity for analytes in the sample to be fractionated, is fixed to an insoluble matrix. The metal ion is immobilized to a solid support matrix via coordinate bonding with chelating agent. The metal ion serves as the characteristic and most essential part of the adsorption center [4]. Protein adsorption is based on coordinate bond formation between the immobilized metal ion and the electron donor group from the protein surface.Most commonly used are the transition-metal ions Cu(II), Ni(II), Zn(II), Co(II), Fe(III), which are electron-pair acceptors and can be considered as Lewis acids. Electron-donor atoms (N, S, O) present in the chelating compounds that are immobilized to the chromatographic support are capable of coordinating metal ions and forming metal chelates, which can be bidentate, tridentate, etc., depending on the number of occupied coordination bonds. The remaining metal coordination sites are normally occupied by water molecules and can be exchanged with suitable electron-donor groups from the protein.In addition to the amino terminus, side chains of certain amino acids bind to the chelated metal, due to their electron-donor atoms. Although many residues, such as Glu, Asp (-ve interaction) Tyr, Cys, His, Arg, Lys, and Met (+ve interaction), can interact with the protein, retention in IMAC is based primarily on the availability of histidyl residues under certain pH and ionic conditions. Free cysteines that could also contribute to binding to chelated metal ions are rarely available in the appropriate, reduced state. However, aromatic side chains of Trp, Phe, and Tyr appear to contribute to retention, if they are in the vicinity of accessible histidine residues [8]. Furthermore, the systematic investigation by E. Sulkowski using model homologous series of proteins showed that histidine, a member of Porath's triad (histidine, tryptophan, and cys...

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“…In the beginning, the ligand was immobilized onto agarose beads and incorporated in an agarose gel [3]. Later, ligands coupled to soluble polymers [4] and monomeric polymerizable derivatives of the ligands that could be copolymerized in a polyacrylamide gel [5] were developed in order to obtain a more homogeneous ligand distribution in the gel. It was shown with proteins such as ribonucleases A and B, cytochromes c and chymotrypsin that separation in IMAGE is based on the same general principles as established by Sulkowski [6] for IMAC.…”

Section: Introductionmentioning

confidence: 99%

Immobilized metal ion affinity gel electrophoresis: Quantification of protein affinity to transition metal chelates

et al. 1996

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This paper describes some recent advances in the methodology of immobilized metal ion affinity gel electrophoresis. Four different ways to incorporate metal chelate ligands in agarose and polyacrylamide-based electrophoresis gels are evaluated, a new polymerizable metal chelating ligand, allyl-2-hydroxy-3-(N,N-dicarboxymethyl)amino-propyl ether, is introduced, and the determination of affinity constants described. The affinities of model proteins (ribonucleases A and B and cytochromes c from different species) for the transition metal chelate iminodiacetic acid-Cu(II) were studied. The results were found to be in agreement with literature data on immobilized metal ion affinity chromatography, and the polymer nature and the different chemistries used influenced the affinity only quantitatively, keeping the basic mechanisms of interaction unchanged.

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Immobilized metal ion affinity electrophoresis

Cited by 16 publications

References 18 publications

Reduction of protein concentration range difference followed by multicolumn fractionation prior to 2‐DE and LC‐MS/MS profiling of serum proteins

Reduction of protein concentration range difference followed by multicolumn fractionation prior to 2‐DE and LC‐MS/MS profiling of serum proteins

Immobilized metal-ion affinity systems for recovery and structure–function studies of proteins at molecular, supramolecular, and cellular levels

Immobilized metal ion affinity gel electrophoresis: Quantification of protein affinity to transition metal chelates

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