Salicylaldehyde-metal-amino acid ternary complex: a new tool for immobilized metal affinity chromatography.
An immobilized salicylaldehyde (sal) was used to build various salicylaldehyde-copper-amino acid (Sal-Cu-AA) complexes which are stable at a range of pH values (2.0-11.0). The complexes were found to bind protein molecules as IMAC resins. Thirteen proteins were examined for their binding to a Sal-Cu-Gly column. The efficacy of the Sal-Cu-AA resin for protein separation were demonstrated by two examples. The first was a new purification process for garlic lectins from garlic crude extract. It seems that in this case the Sal-Cu-AA resins were more selective than IDA resin. The second was immobilization of concanavalin A (Con A) on the resin and using the immobilized Con A for affinity chromatography of mannose-rich glycoprotein ovalbumin. The Con A could be later eluted with EDTA or imidazole and the Sal-containing polymer could be recharged again for further use.
Leibler
D
D
Department of Biophysics, Weizmann Institute of Science, Rehovot, Israel.
Rabinkov
A
A
Wilchek
M
M
D000447:Aldehydes / Q000737:chemistry*; D000596:Amino Acids / Q000737:chemistry*; D000818:Animals; D013437:Carbon-Sulfur Lyases / Q000737:chemistry; D002645:Chickens; D002846:Chromatography, Affinity / Q000379:methods; D003208:Concanavalin A / Q000302:isolation & purification; D003300:Copper / Q000737:chemistry*; D005737:Garlic / Q000737:chemistry; D006639:Histidine / Q000032:analysis; D037102:Lectins / Q000737:chemistry / Q000302:isolation & purification; D010047:Ovalbumin / Q000302:isolation & purification; D037121:Plant Lectins; D010940:Plant Proteins / Q000302:isolation & purification; D010946:Plants, Medicinal; D012116:Resins, Plant / Q000737:chemistry; D012259:Ribonuclease, Pancreatic / Q000737:chemistry
D000447:Aldehydes; D000596:Amino Acids; D037102:Lectins; D037121:Plant Lectins; D010940:Plant Proteins; D012116:Resins, Plant; D003208:Concanavalin A; C013243:salicylaldehyde; D006639:Histidine; D003300:Copper; D010047:Ovalbumin; D012259:Ribonuclease, Pancreatic; D013437:Carbon-Sulfur Lyases; C019243:alliin lyase