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Metalloenzyme

metalloenzyme

(Science: enzyme) An enzyme that contains a bound metal ion as part of its structure. The metal may be required for enzymic activity, either participating directly in catalysis or stabilising the active conformation of the protein.


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experiences with ni-affinity protein purification??

... I presume. Imidazole, especially 250 mM, can change the pH of your buffer. Imidazole can also carry in contaminants like zinc. If you have a metalloenzyme inhibited by zinc, that could contribute to loss of activity in the presence of imidazole which wouldn't necessarily be due to imidazole. ...

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by blcr11
Sun Jun 29, 2008 12:03 am
 
Forum: Molecular Biology
Topic: experiences with ni-affinity protein purification??
Replies: 2
Views: 1451

inactive enzyme after purification with Ni-TED

... Before the purification with Ni-TED the enzyme is active. After the purification there is no activity left. What can be the reason? It is a metalloenzyme and needs Mn -ions for its activity. Could it be that the imidazole in the buffers while purification is the reason? How can I then reactivate ...

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by piefke
Fri May 02, 2008 2:58 pm
 
Forum: Molecular Biology
Topic: inactive enzyme after purification with Ni-TED
Replies: 3
Views: 642


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