However, the geometry of the intermediate allows the pre-bound peptide to rebind if the exchange peptide does not succeed in forming a closed complex. DM would be released from the complex once this assumes a collapsed conformer with the

cluster of interactions between the peptide and the MHCII at the N-terminal stabilized. The above model of DM-mediated peptide exchange is consistent with what has been proposed on the basis of molecular dynamics simulation analysis,[23] suggesting a dual role for DM during peptide binding. First, because of the stabilization of the groove in an ‘exchangeable’ Angiogenesis inhibitor form, DM shifts the control of peptide binding from kinetic to thermodynamic. Second, because of the competition by DM for binding to the P1 pocket ‘neighbourhood’, the effective

free energy threshold for peptide binding is increased. Hence, only peptides with a sufficient affinity for binding can compete for the P1 pocket, which in turn also results in DM dissociation. A critical aspect of the ‘compare-exchange’ model is the existence of an MHCII/two-peptide intermediate. Such an intermediate was also proposed for the exchange learn more reaction in the absence of DM. In particular, the two-peptide/one MHC complex has been adopted to explain observations from several groups indicating an accelerated release of a pre-bound peptide either at the cell surface or in vitro in the presence of free peptide.[12, 58-60] Initially it was thought that the effect of accelerated dissociation was specific because only I-Ed binding peptides were able to accelerate the dissociation of the hen egg lysozyme 107-116/I-Ed complex either on the surface of cells or in purified forms in solution, and high-affinity I-Ed binders did not affect the half-life of purified ovalbumin 323–339/I-Ad complexes.[60] There is evidence that peptides that may not feature a high affinity for a given allele can promote release of a peptide bound to that allele.[58] The replacement

reaction accelerated by a second peptide was indicated as push-off, and was experimentally observed in gels first,[59] and subsequently in solution.[12] In Ureohydrolase particular, the action of a push-off peptide, dynorphin A (dynA-[1-13]) was examined on the dissociation kinetics of the PCC-(89–104)/I-Ek complex. Kinetic analysis, fluorescence resonance energy transfer (FRET), and 19F NMR analysis determined the molecular mechanism of push-off. The results indicated that the first step of push-off is indeed the formation of a two-peptide/one-MHC complex in solution. Although estimates of the relative proportion of the two-peptide/MHCII complex were low in those studies, (1·0–0·1%), these complexes were preferentially associated with the ‘open’ conformer of the pMHCII complex during PAGE analysis.