The first time I learned about structural biology was when I took Comprehensive Biochemistry I during my senior year at University of Maryland, Baltimore County. What was intriguing was how quickly I learned the concepts, and how easy it was for me to see conformational changes in three-dimensional space. I have always been a visual-oriented person, and until senior year of college I found that it was hard to find areas of research in biochemistry that utilize a person’s ability to analyze images and structures, until I discovered structural biology. I took this newfound discovery one step further by enrolling in a senior seminar course on structural biology, something I was apprehensive about because I did not need the course to graduate,
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I found that hS100A7 and its analogs are highly up-regulated in psoriatic keratinocytes, and there is evidence of hS100A7 being up-regulated in some invasive carcinomas but further studies are needed before a definite conclusion can be made. I conducted the purification of the protein under the guidance of Dr. Raquel Ruiz and involved expression tests at different concentrations of the inducing agent IPTG, large-scale growth of BL21-DE3 E. coli cells that over-express hS100A7, utilizing fast protein liquid chromatography (FPLC) with immobilized metal ion affinity and size exclusion columns, and ultimately using the purified protein product in setting up crystallization screens. I was unable to obtain any crystals of hS100A7 due to the usage of the protease inhibitor cocktail AEBSF after cell lysis, which is known to interfere with protein crystallization. The next steps after storage of the frozen protein are to remove any endotoxins in the protein solution in order to be used in in vivo drug …show more content…
Tissue transglutaminase is a cytosolic protein primarily responsible for transamidation reactions of peptide substrates, and TG2 malfunction contributes to celiac disease, cataract formation, and neurodegenerative disorders. Development of TG2 inhibitors may also be an effective route for chemotherapy, because results have suggested that TG2 induces epithelial mesenchymal transition and other stem cell characteristics in cancer cells. Expression and purification of TG2 is procedurally similar to hS100A7 and I am currently in the process of purifying the protein using FPLC under the supervision of Dr. Ruiz, as well as optimizing the purification protocol to obtain higher yields of protein in future purifications. My projects as a lab technician have given me the opportunity to participate in research that involves collaborations between different labs, as opposed to independent research in an academic setting like with my first projects at
I found that hS100A7 and its analogs are highly up-regulated in psoriatic keratinocytes, and there is evidence of hS100A7 being up-regulated in some invasive carcinomas but further studies are needed before a definite conclusion can be made. I conducted the purification of the protein under the guidance of Dr. Raquel Ruiz and involved expression tests at different concentrations of the inducing agent IPTG, large-scale growth of BL21-DE3 E. coli cells that over-express hS100A7, utilizing fast protein liquid chromatography (FPLC) with immobilized metal ion affinity and size exclusion columns, and ultimately using the purified protein product in setting up crystallization screens. I was unable to obtain any crystals of hS100A7 due to the usage of the protease inhibitor cocktail AEBSF after cell lysis, which is known to interfere with protein crystallization. The next steps after storage of the frozen protein are to remove any endotoxins in the protein solution in order to be used in in vivo drug …show more content…
Tissue transglutaminase is a cytosolic protein primarily responsible for transamidation reactions of peptide substrates, and TG2 malfunction contributes to celiac disease, cataract formation, and neurodegenerative disorders. Development of TG2 inhibitors may also be an effective route for chemotherapy, because results have suggested that TG2 induces epithelial mesenchymal transition and other stem cell characteristics in cancer cells. Expression and purification of TG2 is procedurally similar to hS100A7 and I am currently in the process of purifying the protein using FPLC under the supervision of Dr. Ruiz, as well as optimizing the purification protocol to obtain higher yields of protein in future purifications. My projects as a lab technician have given me the opportunity to participate in research that involves collaborations between different labs, as opposed to independent research in an academic setting like with my first projects at