{"id":4104,"date":"2021-04-19T12:14:44","date_gmt":"2021-04-19T16:14:44","guid":{"rendered":"http:\/\/sites.miamioh.edu\/undergraduate-research-forum\/?p=4104"},"modified":"2021-04-20T11:49:48","modified_gmt":"2021-04-20T15:49:48","slug":"a30-p-structural-determination-of-alr1300-in-nostoc-sp-pcc-7120-functional-insights-to-pentapeptide-repeat-proteins","status":"publish","type":"post","link":"https:\/\/sites.miamioh.edu\/undergraduate-research-forum\/a30-p-structural-determination-of-alr1300-in-nostoc-sp-pcc-7120-functional-insights-to-pentapeptide-repeat-proteins\/","title":{"rendered":"A30-P: Structural Determination of Alr1300 in Nostoc sp. PCC 7120: Functional Insights to Pentapeptide Repeat Proteins"},"content":{"rendered":"\n

Alr1300 is a protein of an unknown structure and function in cyanobacteria, Nostoc sp. PCC 71210. This protein is part of the same gene cluster as alr1298 which is from the pentapeptide class of proteins which is a common structure with an unknown function in cyanobacteria. Since the function of proteins in gene clusters tend to be similar, by studying this protein\u2019s structure, more insight can be gained on the biochemical function of pentapeptide repeat proteins and alr1298. Vectors for the native and mutant proteins were constructed. The protein was overexpressed in E. coli, protein purification by affinity chromatography, and structural analysis by X-ray crystallography and NMR. Structural determination of alr1300 may lead to determination of the function of alr1298 and subsequently other PRPs. Alr1300 was observed to form a dimer and C84S-alr1300 was shown in an SDS Page gel to break this dimer. The gels also show the protein was successfully purified by the purification process. Preliminary crystals were observed in crystal trials with C84S-alr1300 and Hampton Research kit HR2-110 crystal screen number 6, however the crystals were later lost. The NMR showed peaks that indicated a disordered part of the protein or aggregated protein. The dimer is likely held together primarily by a disulfide bond by the cysteines. Future work will include continuing to set up crystal trials and continue to optimize NMR conditions. Also, by conducting in situ proteolysis to remove the disordered section of the protein to hopefully improve the X-ray crystallography and NMR results. I plan to attend graduate school for biophysics and this research has given me experience with problem solving in a research setting as well as learning some common techniques.<\/p>\n\n\n\n

Authors: <\/strong>Kristen Rose McGinnis, Roujing Zhang, Shibani Dutta Chowdhury, Shuisong Ni, Michael A Kennedy<\/p>\n\n\n\n

Faculty Advisor:<\/strong> Michael A Kennedy, Chemistry and Biochemistry<\/p>\n\n\n\n

Graduate Student Advisor: <\/strong>Roujing Zhang, Chemistry and Biochemistry<\/p>\n\n\n\n