Date of Last Revision
2023-05-02 14:15:41
Major
Chemistry - Biochemistry
Degree Name
Bachelor of Science
Date of Expected Graduation
Spring 2015
Abstract
SMRT/HDAC Associated Repressor Protein interacts with the long noncoding RNA, produced by SRA, by binding at the RRMs. Three projects were done to prepare the truncated proteins for use in 2D and 3D NMR experiments. The first focuses on RRM 3 and its optimization during the purification process. The second focuses on RRM 2-4, which was found to be missing two alpha helices that may be important for protein stability. These helices can also interact with RRM 3 as well due to the tight association of RRMs 3 and 4. The two step PCR extension of RRM 2-4 was assumed to work based on sequencing and agarose gels. The final project focused on serine to cysteine mutations of RRM 2 to allow site directed spin labeling for use in NMR PRE experiments. Both mutants developed in this project do not affect the protein structure or the residues surrounding the mutation based on wild type and mutant HSQC spectra. The spectra of the spin labeled samples does show broadening some resonances, but may need reductant removed from the buffer after the labeling reaction.
Research Sponsor
Dr. Thomas Leeper
First Reader
Dr. Michael Konopka
Second Reader
Dr. Leah Shriver
Recommended Citation
Christie, Shaun M., "SHARP RNA Recognition Motif Optimizations, Extensions, and Mutations for Use in 2D and 3D NMR Experiments" (2015). Williams Honors College, Honors Research Projects. 61.
https://ideaexchange.uakron.edu/honors_research_projects/61
Included in
Amino Acids, Peptides, and Proteins Commons, Biochemistry Commons, Other Chemistry Commons