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inverted inner membrane vesicle - (Oct/21/2006 )

Hello, I am working now with inverted inner membrane vesicle (IIMV). I prepare IIMV from E. coli cells. I want to insert a kind of purified membrane protein into this vesicle, but unfortunately I don't have a good result until now. I tried to stirring the mixture of purified membrane protein and IIMV o/n, but the membrane protein that I inserted was inactive. I hope someone who have a lot of experiences with IIMV can give me help.

-prihardi-

QUOTE (prihardi @ Oct 21 2006, 09:09 AM)
Hello, I am working now with inverted inner membrane vesicle (IIMV). I prepare IIMV from E. coli cells. I want to insert a kind of purified membrane protein into this vesicle, but unfortunately I don't have a good result until now. I tried to stirring the mixture of purified membrane protein and IIMV o/n, but the membrane protein that I inserted was inactive. I hope someone who have a lot of experiences with IIMV can give me help.



Is the membrane orientation critical in your study? Can you use recostituted liposome instead? You mix your memebrane protein with detergent and lipids, then dialyse away the detergent.

-genehunter-1-

Thank you very much for your assist.

Yes, the membrane orientation is very critical in my study. Because, my enzyme is active when the other enzymes on the inner membrane is active, too. I may use the proteoliposome for this purpose, but the activity of the enzyme is very low.

Best regard.


QUOTE
Is the membrane orientation critical in your study? Can you use recostituted liposome instead? You mix your memebrane protein with detergent and lipids, then dialyse away the detergent.

-prihardi-