Immobilized metal-ion affinity chromatography: imidazole proton pump and chromatographic sequelae. II. Chromatographic sequelae.
Isocratic elution with imidazole of some protein models from a chelating gel, CSFF-IDA-M(II), resulted in their desorption owing to the low pH(6-->4) of the mobile, phase rather than to the imidazole itself (imidazole-generated fall in pH; proton pump). Gradient elution with imidazole was best accomplished when the chelating gel was initially converted into its imidazole complex, CSFF-IDA-M(II)Im. The exploitation of the imidazole-generated proton pump in the IMAC of proteins may enhance the versatility of this type of chromatography.
Sulkowski
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Roswell Park Cancer Institute, Department of Molecular and Cellular Biology, Buffalo, New York 14263, USA.
D000818:Animals; D002417:Cattle; D002614:Chelating Agents / Q000737:chemistry*; D002846:Chromatography, Affinity / Q000379:methods*; D003300:Copper; D003850:Deoxyribonuclease I / Q000302:isolation & purification; D004285:Dogs; D019286:Endopeptidase K / Q000302:isolation & purification; D006454:Hemoglobins / Q000302:isolation & purification; D006801:Humans; D007093:Imidazoles / Q000737:chemistry*; D007098:Imino Acids / Q000737:chemistry*; D007781:Lactoferrin / Q000302:isolation & purification; D008670:Metals; D009211:Myoglobin / Q000302:isolation & purification; D009532:Nickel; D011506:Proteins / Q000302:isolation & purification*; D017494:Proton Pumps; D012259:Ribonuclease, Pancreatic / Q000302:isolation & purification; D015032:Zinc
D002614:Chelating Agents; D006454:Hemoglobins; D007093:Imidazoles; D007098:Imino Acids; D008670:Metals; D009211:Myoglobin; D011506:Proteins; D017494:Proton Pumps; D003300:Copper; C029899:imidazole; D009532:Nickel; D003850:Deoxyribonuclease I; D012259:Ribonuclease, Pancreatic; D007781:Lactoferrin; D019286:Endopeptidase K; D015032:Zinc; C008109:iminodiacetic acid