Metal chelate affinity chromatography, a new approach to protein fractionation

Porath, Jerker; Carlsson, Jan; Olsson, Ingmar; Belfrage, Greta

doi:10.1038/258598a0

Cited by 1,950 publications

(962 citation statements)

References 2 publications

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“…Ce vecteur a été sélectionné car il a montré une bonne efficacité pour délivrer des petits acides nucléiques [7] et des protéines [8] dans le cytosol. Pour fixer la protéine au vecteur, nous avons utilisé l'affinité de l'histidine pour l'ion nickel (Ni 2+ ) immobilisé avec un groupement nitrilotriacétate (NTA) [9], une solution pertinente pour au moins trois raisons. Premièrement, de nombreuses protéines recombinantes sont équipées d'une étiquette polyhistidine (his) afin de pouvoir les purifier par chromatographie d'affinité.…”

Section: Organisation Supramoléculaire De Protéinesunclassified

Amélioration du transfert cytosolique de protéines grâce à leur organisation dans un système de délivrance

Zuber¹

2016

Med Sci (Paris)

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Section: Organisation Supramoléculaire De Protéinesunclassified

Amélioration du transfert cytosolique de protéines grâce à leur organisation dans un système de délivrance

Zuber¹

2016

Med Sci (Paris)

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“…Immobilized metal-ion affinity chromatography (IMAC) was originally introduced by Porath et al [21] for purification of His-tagged proteins and is the most frequently applied technique for phosphopeptide and -protein enrichment nowadays [22]. Thereby, phosphorylated peptides and proteins are bound to the stationary phase by electrostatic interactions with positively charged metal-ions that are on their part bound to the column material via iminodiacetic acid (IDA), nitriloacetic acid (NTA) or Tris-(carboxymethyl)-ethylendiamine (TED) linkers.…”

Section: Detection Of Phosphoproteins In 2-d Gelsmentioning

confidence: 99%

State-of-the-art in phosphoproteomics

2005

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Presently, phosphorylation of proteins is the most studied and best understood PTM. However, the analysis of phosphoproteins and phosphopeptides is still one of the most challenging tasks in contemporary proteome research. Since not every phosphoprotein is accessible by a certain method and identification of the phosphorylated amino acid residue is required in the majority of cases, various strategies for the detection and localization of phosphorylations have been developed. Identification and localization of protein phosphorylations is mostly done by MS nowadays but phosphoproteins and -peptides are often suppressed in comparison to the unphosphorylated species if measured in complex mixtures. Thus, the isolation of pure phosphopeptide samples is a main task. This review gives an overview over the most frequently used methods in isolation and detection of phosphoproteins and -peptides such as specific enrichment or separation strategies as well as the localization of the phosphorylated residues by various mass spectrometric techniques.

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“…6, but the relatively low solubility of 5-Br-PADAP and high cost of Nitro-PAPS still restrict the wide use of these reagents. Potentially, even more sensitive reagents could be the porphyrins, e. g., meso-tetrakis(4-N-trimethylaminophenyl)porphine (TTMAPP), which gives molar extinction coefficients for the copper complex of 4.8610 5 L N mol -1 N cm -1 [65]; however, the first attempt of their…”

Section: New Reagents For Pcrmentioning

confidence: 99%

“…Chelation-in-a-sorbent based chromatographic techniques embrace ligand-exchange chromatography developed by Helfferich in 1961 [4], immobilized metal ion affinity chromatography introduced by Porath in 1975 [5], and silver ion or argentation chromatography introduced by Morris in 1962 [6]. These techniques are now widely used for the separation of polar organic molecules such as amines, amino alcohols and amino acids, phenols, carbohydrates, lipids, unsaturated fatty acids, peptides, proteins, etc.…”

Section: Introductionmentioning

confidence: 99%

Recent developments in the high‐performance chelation ion chromatography of trace metals

Nesterenko¹,

Jones

2007

J of Separation Science

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Review Recent developments in the high-performance chelation ion chromatography of trace metalsThere have been a number of significant developments in the high-performance chelation ion chromatography (HPCIC) of trace metals in recent years. This review focuses on these developments, while giving important information on the fundamental parameters controlling the chelation sorption mechanism, including type of chelating group, stability constants, kinetics, and column temperature. The discussion pays particular attention to the types and properties of efficient chelating stationary phases which have been fabricated for certain groups of metals. The review also describes a number of major improvements in postcolumn reaction detection including the use of the latest reagents and noise reduction strategies to improve sensitivity and reduce LOD. In the final section, an indication of the applicability of HPCIC to a range of complex sample types is given with some key examples and chromatograms using the latest high-efficiency chelating phases.

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Metal chelate affinity chromatography, a new approach to protein fractionation

Cited by 1,950 publications

References 2 publications

Amélioration du transfert cytosolique de protéines grâce à leur organisation dans un système de délivrance

Amélioration du transfert cytosolique de protéines grâce à leur organisation dans un système de délivrance

State-of-the-art in phosphoproteomics

Recent developments in the high‐performance chelation ion chromatography of trace metals

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