As expected, Bcl 2 only in cells, which detects the human KRN 633 KRN633 protein. Especially if also Bax in each of the paired responses Co he executed with Bcl 2 as well as from reactions containing tBID to falls. Zus Tzlich  in the presence of tBid. Was added sufficient for tBid to adjust the conformation of most or all of the Bcl 2 on the membrane, the result shows that the conformation ver Changed Bcl 2 binds to Bax. Is from the moment Bax incorporated as integral membrane protein helices 5, 6 and 9 in the membrane, Close s assume that the conformation ver Bcl 2 changed with the membrane as an integral member Bax binds.
Under the same conditions, Rolipram Bax eluted from the S Gelfiltration molecules as small oligomer 40 50 kDa instead of the 4200 kDa oligomers found without Bcl second Thus inhibiting the binding of Bcl ver 2 conformation S changing the form of integral membrane proteins Bax oligomerization of Bax. Several observations suggest that only binds tBid fa Transient Bcl 2 is to activate it, and not be trapped by the second Bcl First, tBid can with Bcl 2 at concentrations 420 times h Since we are linked to the conformational Use change. Secondly, if tBid mitochondrial Bcl contain 2 and mitochondria were pelleted added tBid remained in the supernatant. After all, Bcl 2 binds to and Bid tBID CHAPS, but not in a non-ionic detergent is known that the conformation of the two members of the Bcl ver Change form active and capable of inserting in a membrane mimics.
It is likely that the conformational tBid alone can drive Change in Bcl 2, because the conformation Induced change of Bcl 2 tBid through, even if not added Bax. Au Addition, the dependence Dependence of Identical change in the conformation of tBid Bcl 2 is essentially when the above shot Bax was present at. Similar results were also observed when Bim peptide was used to induce conformational Change of Bcl second However, it is possible to change that formally activated Bax can also cause these conformational Change of Bcl second If the ver MODIFIED conformation Bcl 2 inhibits Bax integral membrane proteins Form, the amount of the two proteins Should in mitochondria hen to increased in a coordinated way, And the report would show them to their pc Stoichiometry complexes Bax inhibited the mitochondrial membranes.
If we the H Integrated Bax he difficult in membranes of the amount of conformationally ver Changed Bcl 2 as compared to addition of different amounts tBid, there was a good match with almost equal amounts of the two species. Thus, in response to 500 nM and 20 nM Bax Bcl 2 when tBid added to 1 nM, the extent the Ver change conformation Bcl 2 and Bax was integral membrane exhibit very similarB8 respectively and 6 nm. Ht as tBid erh Was 4 or 8 nM, cytochrome c release from isolated mitochondria from cells detected bcl 2 and exceeded the amount of integrated Bax conformation ver Changed Bcl second Although this result suggests that the conformation ver Changed Bcl 2 inhibits Bax via an integral membrane protein, beat the gel filtration data in Figure 3 that the mechanism is not so simple to connect. W During elution