A clear difference between the two sequences is the selleck bio number Inhibitors,Modulators,Libraries of residues present between two YPs. Since F122 constitutes Pocket 2 and interacts with the aromatic side chain of the corresponding sec ond YP in the Alix ABS peptide, mutation of F122 seems favored or disfavored for interaction depending 10. 7. On the other hand, regardless of the disruption of inter helix interaction, ALG 2GF122 gave only a small increase in the angle in the Ca2 bound form structure, and the dis tance between the Ca atoms of Y124 and T160 was sig nificantly Inhibitors,Modulators,Libraries decreased in ALG 2GF122. Consequently, the change in the relative spatial positions of a5 and the following loop may allow the side chains of A122, Y124 and adjacent R125 to place at more appropriate positions in the hydrophobic pockets of ALG 2F122A to interact with the Alix ABS peptide.
To corroborate the above hypothetical mechanism of Inhibitors,Modulators,Libraries enhanced binding of ALG 2F122A to Alix, we attempted co crystallization of des3 20ALG 2F122A with the Alix ABS peptide but could not obtain good crystals. The nature of N terminal residues of ALG 2 influences qualities of crystals. In the present study, we used des3 20ALG 2F122A because X ray diffraction reso lutions were better in des3 20ALG 2 than in des3 23ALG 2 in the Ca2 bound forms in our previous study. It would Inhibitors,Modulators,Libraries be worthwhile, however, to test des3 23ALG 2F122A for crystal preparations of the complex. ALG 2 interacts with various proteins with Pro rich regions. Previously, we classified ALG 2 interacting proteins into two groups based on ability of binding to ALG 2GF122.
Alix type proteins such as TSG101, annexin A7 and annexin A11 contain a consensus sequence of PPYPX3 5YP similar to the core motif of Alix. On the other hand, ALG 2GF122 retains bind ing ability to non Alix type Inhibitors,Modulators,Libraries proteins such as Sec31A and phospholipid scramblase 3 that possess a common sequence of PXPGF but on spatial positions of the concerned Tyr residues in the target proteins. Recently, mucolipin 1 has been shown to Ca2 dependently interact with ALG 2 but not with ALG 2GF122. The predicted binding site does not contain any YP or PXPGF motif, but charged residues as well as hydropho bic residues were shown to be important for interac tions. Thus, mucolipin 1 may be recognized by a surface different from that for binding to Pro rich target pro teins.
Ask1 and Raf 1, which are also known to interact with ALG 2, do not possess conspicuous Pro rich regions e-book either, and the ALG 2 binding site has not been reported yet. It would be interesting to see whether F122A and other amino acid substituted mutants of Pocket 1 and Pocket 2 present a different binding profile between Pro rich type and non Pro rich type ALG 2 interacting proteins. The biological significance of the occurrence of ALG 2GF122 is not known. ALG 2 forms a dimer and each molecule of the dimer has capacity of Alix binding.