Chelated mercury as a ligand in immobilized metal ion affinity chromatography of proteins

Gelūnait, Lina; Lukša, Virginijus; Sūdžiuvien, Ona; Bumelis, Vladas Algirdas; Pesliakas, Henrikas

doi:10.1016/s0021-9673(00)00886-4

Cited by 13 publications

(8 citation statements)

References 48 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…37,49,[57][58][59][60][61][62][63] A literatura mostra que, além destas aplicações, IMAC foi utilizada para separação de diferentes tipos de células microbianas, uma vez que estas, quando possuíam resíduos de histidina acessíveis, apresentavam a capacidade de serem adsorvidas em uma coluna monolítica de estrutura macroporosa composta por criogéis de poliacrilamida com IDA-Cu 2+ imobilizado. 65 Proteínas que contêm naturalmente resíduos de histidinas acessíveis (recombinantes ou não) e apresentam importância comercial no campo terapêutico e diagnóstico, como proteínas do soro humano, 29 imunoglobulinas, 31,35,38,[66][67][68][69][70][71] interferon, 72 proteína C humana, 73,74 fator IX 73,75 e hormônio de crescimento humano, 76 foram purificadas por IMAC utilizando os metais de transição Cu 2+ , Ni 2+ , Zn 2+ , Fe 3+ e Co 2+ imobilizados.…”

Section: Exemplos De Aplicações De Imacunclassified

Cromatografia de afinidade por íons metálicos imobilizados (IMAC) de biomoléculas: aspectos fundamentais e aplicações tecnológicas

Bresolin¹,

Miranda²,

Bueno³

2009

Quím. Nova

View full text Add to dashboard Cite

Recebido em 16/7/08; aceito em 16/12/08; publicado na web em 11/5/09 IMMOBILIZED METAL-ION AFFINITY CHROMATOGRAPHY (IMAC) OF BIOMOLECULES: FUNDAMENTAL ASPECTS AND TECHNOLOGICAL APPLICATIONS. Immobilized Metal Ion Affinity Cromatography -IMAC -is a group-specific based adsorption applied to the purification and structure-function studies of proteins and nucleic acids. The adsorption is based on coordination between a metal ion chelated on the surface of a solid matrix and electron donor groups at the surface of the biomolecule. IMAC is a highly selective, low cost, and easily scaled-up technique being used in research and commercial operations. A separation process can be designed for a specific molecule by just selecting an appropriate metal ion, chelating agent, and operational conditions such as pH, ionic strength, and buffer type.Keywords: immobilized metal-ion affinity chromatography; purification; biomolecules. INTRODUÇÃOOs princípios fundamentais da afinidade de biomoléculas por íons metálicos são conhecidos desde o início do século passado. Em 1974, Everson e Parker demonstraram que íons metálicos presentes em metaloproteínas são os principais responsáveis pela adsorção dessas em resinas contendo quelantes imobilizados e exploraram esta afinidade na separação deste tipo de proteínas. 1 A técnica proposta por Everson e Parker popularizou-se com o trabalho publicado por Porath e colaboradores em 1975, quando os autores introduziram o termo cromatografia de afinidade por íons metálicos imobilizados (Immobilized Metal-Ion Affinity Chromatography -IMAC). 2 IMAC explora a interação entre espécies doadoras de elétrons presentes na superfície de biomoléculas em solução e íons metálicos quelatados imobilizados em um suporte sólido. Inúmeras são as aplicações analíticas, preparativas e industriais de IMAC, como a separação e purificação de diferentes biomoléculas (peptídeos, proteínas e ácidos nucléicos), a separação de células a partir de extratos biológicos e estudos de estrutura-função de proteínas.Inicialmente a técnica de IMAC foi extensamente utilizada para purificação de biomoléculas contendo naturalmente grupos doadores de elétrons em resíduos de aminoácidos expostos na superfície (tais como o anel imidazol de histidina), os quais são primariamente responsáveis pela interação biomolécula-íon metálico. Com o advento da tecnologia do DNA recombinante, foi possível a incorporação de caudas (tag) em proteínas que não contêm naturalmente espécies doadoras de elétrons, tal como a fusão de sequência de seis histidinas na porção C ou N-terminal da proteína alvo, conferindo à mesma a possibilidade de purificação por IMAC. 3,4 Apesar de na literatura existirem excelentes artigos de revisão em inglês sobre IMAC 5-14 , nesta revisão, devido à intensa atividade neste campo, pretende-se abordar aspectos fundamentais, aplicações tecnológicas (escala de laboratório e industrial) e científicas com ênfase na purificação de proteínas nativas e recombinantes (com cauda de poli(histidina)). Dentre os aspectos fundamentais do mé...

show abstract

Section: Exemplos De Aplicações De Imacunclassified

Cromatografia de afinidade por íons metálicos imobilizados (IMAC) de biomoléculas: aspectos fundamentais e aplicações tecnológicas

Bresolin¹,

Miranda²,

Bueno³

2009

Quím. Nova

View full text Add to dashboard Cite

show abstract

“…The rapid development of biotechnology and biomedicine requires more reliable and efficient separation techniques for the isolation and purification of biomolecules such as proteins, enzymes, nucleic acids and hormones. Immobilized metal ion affinity chromatography (IMAC) is an efficient method for purification of biomolecules in both analytical and large‐scale modes 1–4. It is based on the selective interaction of immobilized metal ions with proteins through their surface‐exposed amino acid residues such as histidine, cysteine and tryptophan 5–8.…”

Section: Introductionmentioning

confidence: 99%

Synthesis of monodisperse nonporous crosslinked poly(glycidyl methacrylate) particles with metal affinity ligands for protein adsorption

Guan

Liu

2005

Polymer International

View full text Add to dashboard Cite

Monodisperse nonporous crosslinked poly(glycidyl methacrylate) (PGMA) particles with immobilized metal affinity ligands were prepared for selective recovery of proteins. The PGMA particles, with an average size of 2.2 µm, were prepared by a simple dispersion polymerization of glycidyl methacrylate (GMA) and ethylene glycol dimethacrylate (EGDMA). The particles were characterized by scanning electron microscopy (SEM) and Fourier-transform infrared spectroscopy (FTIR). The epoxy groups of the particles were modified with the metal chelating agent iminodiacetic acid (IDA), which forms metal-IDA chelates at the active sites. After charging with copper ions, the particles were used to recover a model protein, bovine hemoglobin (BHb), in a batchwise manner. The particles had the adsorption capacity of 218.7 mg g −1 with little nonspecific adsorption. The adsorption behavior could be described with the Langmuir equation. The effect of pH on the adsorption was also studied. Regeneration of the metal-chelated particles was easily performed with 50 mmol L −1 ethylenediaminetetraacetic acid (EDTA), followed by washing with water and reloading with Cu 2+ . The particles could be very useful as an affinity separation adsorbent.

show abstract

“…Mrksich and coworkers quantitatively evaluated the effect of three different inhibitors of the non‐receptor tyrosine kinase, c‐Src by immobilizing a 9‐mer peptide on a gold surface, thus shedding light on the dynamics of enzyme–substrate interactions and the applications in drug discovery . Moreover, immobilized metal‐ion affinity chromatography (IMAC) is often used for the analysis of analytes and the purification of proteins …”

Section: Methodsmentioning

confidence: 99%

A Facile Method for Detection of Substituted Salicylic Acids Using Pyrenesulfonamide‐Terminated Self‐Assembled Monolayers on Silicon Oxide Surfaces

Han

Kumar

Choi

et al. 2016

Bulletin Korean Chem Soc

View full text Add to dashboard Cite

Self-assembled monolayers (SAMs) of alkylsilanes (e.g., alkyltrialkoxysilanes and alkyltrichlorosilanes) on Si/SiO 2 surfaces have received a great deal of attention in a variety of research areas potentially because the SAMs improve the fundamental understanding of interfacial phenomena and develop many technological applications, such as fieldeffect transistor, catalysis, microarray, non-biofouling coating, and (bio)analytical devices. 1 In the field of chemo/biosensors, immobilization of receptors on surfaces enables the functional and structural understanding of the molecular interactions between receptors and analytes and usually provides rapid response times, easy-to-use testing procedures, and high potential for the reuse of materials. 2 For example, a number of protein functions were successfully investigated through the immobilization of over 10 000 proteins on a Si/SiO 2 surface. 3 Mrksich and coworkers quantitatively evaluated the effect of three different inhibitors of the non-receptor tyrosine kinase, c-Src by immobilizing a 9-mer peptide on a gold surface, thus shedding light on the dynamics of enzyme-substrate interactions and the applications in drug discovery. 4 Moreover, immobilized metal-ion affinity chromatography (IMAC) is often used for the analysis of analytes and the purification of proteins. 5 Salicylic acids (o-hydroxybenzoic acids) play a pivotal role in the metabolism and physiology of living organisms. 6 They are also widely used in pharmaceuticals, cosmetics, medical diagnosis, food technology, and environmental monitoring. 7 For example, in plants, salicylic acid is involved in the regulation of defenses against pathogens and insects and affects a number of physiological processes such as fruit yield, seed germination, protein synthesis, and nutrient uptake. 8 In addition, salicylic acid is an aromatic organic compound, which may cause allergic contact dermatitis, one of the most prevalent forms of immunotoxicity found in humans. 9 Consequently, monitoring and quantita-tive determination of salicylic acids in aqueous solutions is important. 2,10 Moreover, for detailed studies on their function and physiological properties and effects, it is necessary to be able to selectively recognize each derivative.Recently, we have successfully demonstrated the molecular recognition between salicylic acids and pyrene derivatives in solution by fluorescence measurement. Briefly, selective recognition was achieved using intermolecular interactions, including π-π interactions and multi-hydrogen bonds, and intramolecular hydrogen bonding between the phenolic O-H group and the adjacent C O group. 11 More interestingly, the association constant for the binding event was affected by the substituents of both the pyrene and salicylic acid moieties. For example, pyrene-1-butylsulfonamide has different association constant values with various salicylic acid derivatives: 2.83 × 10 4 M −1 for 3,5dinitrosalicylic acid, 9.07 × 10 3 M −1 for 5-nitirosalicylic acid, and 5.32 × 10 3 M −1 for 5-iodosalicylic ac...

show abstract

Chelated mercury as a ligand in immobilized metal ion affinity chromatography of proteins

Cited by 13 publications

References 48 publications

Cromatografia de afinidade por íons metálicos imobilizados (IMAC) de biomoléculas: aspectos fundamentais e aplicações tecnológicas

Cromatografia de afinidade por íons metálicos imobilizados (IMAC) de biomoléculas: aspectos fundamentais e aplicações tecnológicas

Synthesis of monodisperse nonporous crosslinked poly(glycidyl methacrylate) particles with metal affinity ligands for protein adsorption

A Facile Method for Detection of Substituted Salicylic Acids Using Pyrenesulfonamide‐Terminated Self‐Assembled Monolayers on Silicon Oxide Surfaces

Contact Info

Product

Resources

About