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Avoiding PTM Actylation of rProtein

PTM

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5 replies to this topic

#1 Pangea

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Posted 03 January 2013 - 10:00 AM

How can İ prevent acylation (Lys) of my recombinant protein in E. coli or Mammalian cells?

#2 HOYAJM

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Posted 07 January 2013 - 08:55 AM

Could you treat the cells with an acyltransferase inhibitor? They are available commericially.

#3 Pangea

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Posted 07 January 2013 - 10:09 AM

This option also came in my mind but to knockout the gene would be Lethal i guess. In mammalian maybe one could use stable cell lines transfected with shRNA, or?

#4 Pangea

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Posted 07 January 2013 - 10:09 AM

Is there a way to screen a inhibitor collection?

#5 HOYAJM

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Posted 07 January 2013 - 11:48 AM

Im not quite following. You fear that if you use an acyltransferase inhibitor you will "knock out" the gene/protein function? If you want to prevent acylation, these inhibitors seem like the logical choice. It will inhibit acylation of all proteins, but there will be a recommended dose to use that should maintain a majority of protein functions.

#6 Pangea

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Posted 07 January 2013 - 12:28 PM

No of course inhibition will be happing on the protein level. I mean cutting out the gene will be lethal. But thanks for sharing your knowledge.





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