TY - GEN A1 - Sowemimo, Oluwakemi A1 - Borcherds, Wade A1 - Knox-Brown, Patrick A1 - Rindfleisch, Tobias A1 - Thalhammer, Anja A1 - Daughdrill, Gary T1 - Evolution of Transient Helicity and Disorder in Late Embryogenesis Abundant Protein COR15A T2 - Biophysical journal N2 - Cold regulated protein 15A (COR15A) is a nuclear encoded, intrinsically disordered protein that is found in Arabidopsis thaliana. It belongs to the Late Embryogenesis Abundant (LEA) family of proteins and is responsible for increased freezing tolerance in plants. COR15A is intrinsically disordered in dilute solutions and adopts a helical structure upon dehydration or in the presence of co-solutes such as TFE and ethylene glycol. This helical structure is thought to be important for protecting plants from dehydration induced by freezing. Multiple protein sequence alignments revealed the presence of several conserved glycine residues that we hypothesize keeps COR15A from becoming helical in dilute solutions. Using AGADIR, the change in helical content of COR15A when these conserved glycine residues were mutated to alanine residues was predicted. Based on the predictions, glycine to alanine mutants were made at position 68, and 54,68,81, and 84. Labeled samples of wildtype COR15A and mutant proteins were purified and NMR experiments were performed to examine any structural changes induced by the mutations. To test the effects of dehydration on the structure of COR15A, trifluoroethanol, an alcohol based co solvent that is proposed to induce/stabilize helical structure in peptides was added to the NMR samples, and the results of the experiment showed an increase in helical content, compared to the samples without TFE. To test the functional differences between wild type and the mutants, liposome leakage assays were performed. The results from these assays suggest the more helical mutants may augment membrane stability. Y1 - 2019 UR - https://publishup.uni-potsdam.de/frontdoor/index/index/docId/50217 SN - 0006-3495 SN - 1542-0086 VL - 116 IS - 3 SP - 473A EP - 473A PB - Cell Press CY - Cambridge ER -