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Thread: freeze clamp as camera?

  1. #1 freeze clamp as camera? 
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    May 2012
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    guys,

    I'd like to discuss this idea (on youtube on the link below) to record which molecule(s) is(are) interacting with which ones when two kind of cells met. So far, in any case, at least one of the partner has to be know. The game then consist to try to identify who is binding to it (blotting, screening, labeling). Most of the time, it required firstly purification of known proteins. Here the idea is toreverse the mapping of cell-cell, cell-protein and protein-protein interaction by freezing, linking them with UV and then sequence the attached parts after isolation to identify the interacting partners. I use freeze clamp to freeze ionic channel so, they can be labeled with photoactive probes.
    My hypothesis is to use photoactivatable probes in the media where are the cells. When they become spatially very close to each other (example during an invasion), at their interfaces, some probes will be trapped. If the surfaces are complementary geometrically, the bond will be stronger (?). When the media is instant frozen, each partner will stay in its exact position at a precise moment of the molecular events happening. Nitrogen freezing enables the fast freezing. So, if the arms of the probe is close enough of each surface molecules, the UV irradiation will bind them together covalently. If the trio (part of cell A, part of cell B and the photoactive linker) are labeled with specific "dyes" or labels (radio-elements?), the all process will be to track any band on gel having the three signals. These trio bands having the label of the three markers will be purify and sequenced (liquid or Mass spec). The obtained sequence enables the identification of the partner in data base. Therefore, no need to know the interacting partners before sequencing them...
    Thank you for your enlightenment

    HB
    .


    link youtube: http://www.youtube.c...h?v=5QMrb-d9NaM


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