Chromatographic purification of a mammalian histidine decarboxylase on charged and non-charged alkyl derivatives of agarose.
Histidine decarboxylase (EC 4.1.1.22) from a mouse mastocytoma has been purified by chromatography on charged and non-charged n-alkyl derivatives of agarose. The former was represented by the coupling product of CNBr-activated agarose and alkylmonoamines (alkylamino-agarose), the latter by the coupling of agarose and alkylglycidyl ehters (alkyl agarose). The choice of fractionation medium was restricted by the enzyme stability; excessively high ionic strength media could not be used. Under the conditions investigated, the best result was obtained with the non-charged ocytl agarose. The enzyme was adsorbed to this gel at a relatively high ionic strength, and on stepwise decrease in ionic stength of the eluting buffer it was desorbed with a total recovery of 80%. There was an approx. 10-fold increase in specific activity. The histidine decarboxylase, thus purified, retained 90-100% of its activity for 10 days or more at 6-8 degrees C. Some general comments on protein fractionation on charged and non-charged alkyl derivatives of agarose are given. The complexity of protein interaction with the charged alkyl derivatives is illustrated by experiments with a colored protein, phycoerythrin.
D000818:Animals; D002262:Carboxy-Lyases / Q000302:isolation & purification*; D002846:Chromatography, Affinity; D006640:Histidine Decarboxylase / Q000302:isolation & purification* / Q000378:metabolism; D012515:Mast-Cell Sarcoma / Q000201:enzymology*; D051379:Mice; D009374:Neoplasms, Experimental / Q000201:enzymology; D012685:Sepharose
D012685:Sepharose; D002262:Carboxy-Lyases; D006640:Histidine Decarboxylase