Next we modeled the possible interaction of AIF and endoG resulting in global energy value 26. 27 for this complex. Lastly, we modeled experimen tally proved interaction of analogues of AIF and endoG, WAH 1 and CPS 6. Resulting global energy was 13. 11. Figure 5 shows parts of modeled docking selleck chemicals Gefitinib complexes described above with focus on visualization of binding Inhibitors,Modulators,Libraries sites. In green color are shown residues predicted by cons PPISP or DISPLAR servers mentioned above that can form binding sites for protein protein and protein DNA interactions. Discussion We clearly show that sequence homology of AIF and AMID is restricted to Ndh conserved domain, that corre sponds to oxidoreductase function and not to apoptotic function of AIF, which was shown to reside in large C ter minal part of AIF sequence, which is totally missing in AMID sequence.
AMID sequence contains, aside from Ndh conserved domain and N terminal part, only very small C terminal part consisting of only several residues and thus it is highly improbable that AMID will act similarly as AIF during apoptosis. We correctly pre dicted that endoG and AIF would localize Inhibitors,Modulators,Libraries to the mito chondria, HSP70 1 and cyclophilin A to the cytoplasm, and DNA topoisomerase II to the nucleus. The cytoplasm was found as the most probable cellular loca tion for AMID. This result is in agreement with previous observations, although some of them described the distribution of AMID in the cytoplasm dif ferently. No recognizable localization signal was Inhibitors,Modulators,Libraries found in the AMID amino acid sequence.
Inhibitors,Modulators,Libraries However, the MLS was detected as the first 48 amino acids of the endoG sequence and as the first 61 amino acids of the AIF sequence. This finding corroborates the many pub lished Inhibitors,Modulators,Libraries observations that these proteins are translocated into the nucleus during apoptosis. Interestingly, the predicted NLS of endoG is located within the MLS, so that once endoG enters the mitochondrion the NLS will be cleaved together with the MLS. NLS was found in sequences of DNA topoisomerase II , which was expected, due to function of this protein. Interestingly, NLS was also found in the sequence of HSP70 1 suggest ing its possible role in nucleus. HSP70 1 was found in nucleoli after the heat shock. The algorithms and TMHMM 2. 0 server also predicted that the sequence of endoG, cyclophilin A, HSP70 1 and DNA topoisomerase II do not contain any transmembrane regions.
However, PSORTII algorithms and TMHMM 2. 0 revealed that AIF and AMID sequences contain one predicted transmem brane region selleck chem thus suggesting that AIF and AMID could be membrane proteins. When apoptosis is induced the AIF protein is spliced probably by calpain I and looses its N terminal part that contains the predicted transmem brane region and AIF is thus released into the intermem brane space and out of mitochondria.