%0 PDF %T Conformational Restriction of the NPF Motif to Target EHD1 and Endocytic Recycling %A KAMENS, ALISSA. %D 2017-06-29T09:04:37.223-04:00 %8 2017-07-07 %R http://localhost/files/bc386w62x %X Abstract: The development of specific inhibitors for the long-loop recycling pathway is critical for the study of vesicle trafficking and its involvement in the spread of metastasized cancer cells. Many of the proteins involved in this pathway are difficult to target due to the large and shallow protein-protein interactions they require to function. EHD1, a C-terminal EH domain-containing protein, is a promising target for this process due to its involvement in surface receptor recycling. In addition, many of its interacting proteins do so through an asparagine-proline-phenylalanine (NPF) motif, which forms a type 1 β-turn in the hydrophobic binding pocket of the EH domain of EHD1 (EHD1-EH). In the work described in this thesis, we have developed inhibitors of EHD1 by synthesizing a cyclic peptide with an NPF sequence. The incorporation of the NPF motif into a backbone macrocyclic peptide appeared to stabilize its naturally occurring β-turn, and the cyclic peptide inhibitors bound EHD1-EH more tightly than the corresponding linear controls. We then developed our best inhibitor of the first design series, cNPF1, into a fluorescent probe to further study EH domain binding and screen for stronger EH domain inhibitors. Our newest series of inhibitors utilize the NPF motif in conjunction with a new synthetic pathway to explore a variety of linkers with a series of binding epitopes. The bis-alkylation of thiol-containing peptides allowed for many linkers to be examined in the context of the same peptide sequence. We search for more beneficial binding conformations, while ideally improving passive cell penetration through the incorporation of a hydrophobic linker. These higher affinity inhibitors will help us gather a better understanding of the effects of EHD1 in the context of cancer cell invasion.; Thesis (Ph.D.)--Tufts University, 2017.; Submitted to the Dept. of Chemistry.; Advisor: Joshua Kritzer.; Committee: David Walt, Clay Bennett, and Robert Hammer.; Keywords: Chemistry, Biochemistry, and Biophysics. %[ 2022-10-11 %9 Text %~ Tufts Digital Library %W Institution