Structural Biology Reflection

Decent Essays
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, …show more content…
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

Related Documents

  • Decent Essays

    Tumor cells often take advantage of these mechanisms by using immunological brakes or checkpoints to escape the immune system. Cytotoxic T -lymphocyte-associated antigen 4 (CTLA-4) and Programmed death 1 (PD-1) are targets for cancer treatment. CTLA-4 is a protein found on surface of T cells that binds to B7 family molecules expressed by dendritic cells through an inhibitory pathway, thus preventing CD28 from binding to B7 (Pardoll 2012). This pathway leads to suppression of T cell activation freeing cancer cells from immunosurveilance. Researchers have developed monoclonal antibodies (drugs) for CTLA-4 to activate the CD28/B7 T cell activation and elicit an immune response against tumors (Alegre & Fallarino 2006).…

    • 1172 Words
    • 5 Pages
    Decent Essays
  • Decent Essays

    Adaptive Immune Response

    • 785 Words
    • 4 Pages

    The hypervariable regions are mutated for production of high affinity antibodies. Immunoglobulin genes undergo a process of somatic hypermutation. Somatic hypermutation is important for the generation of high affinity antibodies, it introduces point mutations at a very high rate into variable regions of the rearranged heavy and light chain genes. This results in mutated immunoglobulin molecules on the surface of the B cells. Somatic recombination involves gene segments of the heavy and light chains’ variable, diversity and joining regions.…

    • 785 Words
    • 4 Pages
    Decent Essays
  • Decent Essays

    T Cells Essay

    • 1316 Words
    • 6 Pages

    Scientists have demonstrated that adaptive immunity intervened by T-cell lymphocytes gives important information and support for implementing and maintaining antitumor response. (citation). a broad tumor ingression by cytotoxic CD8+ T cells was heavily correlated with patient survival and react to therapy. Hence, additional studies have observed that cancer reversion and autoimmunity caused by cytotoxic T lymphocyte-associated antigen 4 blockade in patients with metastatic melanoma. Cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) is an essential immune-regulatory molecule (showed on triggered T cells and a portion of regulatory T cells) able to down-regulate T cell activation.…

    • 1316 Words
    • 6 Pages
    Decent Essays
  • Decent Essays

    Thus far, there are three reports in literature that examine the function of ATF3 in different stromal cells—fibroblasts, pericytes, and tumor-associated macrophages; and I will recapitulate them here. 1) Buganim et.al. reported that ATF3 over-expression in cancer-associated fibroblasts (CAFs) promoted tumor growth of co-injected cancer cells in mice. ATF3 expression in CAFS transcriptionally represses CLDN1 and induces CXCL12 and RGS4 thereby mediating its pro-tumor effect. 2) Xue et.al.…

    • 1213 Words
    • 5 Pages
    Decent Essays
  • Decent Essays

    AKAP9-BRAF in thyroid cancer45 and KIAA1549-BRAF in pilocytic astrocytoma46 have also been reported by others and recently pancreatic acinar cell carcinoma was known to have RAF kinase rearrangements similar to the fusion genes that we identified in prostate cancer but with different tissue specific 5’partner gene27. Similarly “druggable” kinase gene fusion, EML4-ALK47 was identified in 1-5% of non-small cell lung cancer. Based on our experience in using the next generation sequencing technology we believe that transcriptome sequencing is the rational and an unbiased approach for the identification of gene fusions in the ETS negative AA men with prostate cancer44;…

    • 1479 Words
    • 6 Pages
    Decent Essays
  • Decent Essays

    Explain how changes in the following three types of genes can cause cancer ○ Explain the role of Tumor Suppressor Genes when functioning normally. The tumor suppressor gene controls Ras genes and P53, it slows cell division ○ What are the mutations that can occur in those genes? Missense, Nonsense, Deletion, Insertion, Substitution, Inversion, Duplication ■ How do those mutations cause cancer? Chromosome rearrangements: Changes in chromosomes that put one gene next to another, which allows one gene to activate the other Gene duplication: Having extra copies of a gene, which can lead to it making too much of a certain protein ■ Provide 2 common examples Familial retinoblastoma: occurs when a baby inherits from one of its parents a chromosome (number 13) that has its RB locus deleted. The normal Rb protein controls the cell cycle.…

    • 1004 Words
    • 5 Pages
    Decent Essays
  • Decent Essays

    Nanomedicine Essay

    • 1679 Words
    • 7 Pages

    Especially, activated T cells are divided into effector T cell and memory T cell for effective defense system against secondary infection. [2] Cancer, which is classed as representative intractable disease, expresses TAA or mutated protein onto the surface of cancer cell. It also activates…

    • 1679 Words
    • 7 Pages
    Decent Essays
  • Decent Essays

    The main objective of this paper by Ando et al. was to explore the nature of association between the protein MEF/ELF4 and the nucleophosmin, NPM1, along with its implications with regards to leukemogenesis; the mutant version of the latter has been commonly associated with leukemia. MEF/ELF4 is part of a family of transcriptional factors (ETS) responsible for numerous cellular functions, such as genomic stability, DNA repair, and most importantly, the regulation of cell proliferation, differentiation and cell death. In particular, MEF/ELF4 is expressed in hematopoietic cells in order to control their movement through the eukaryotic cell cycle, thus classifying it as a potential oncogene. Previous studies have demonstrated that the human HDM2 promoter is a target of the MEF transcriptional factor and the overexpression of MEF led to an increase in the Mdm2 protein (mice version of the human Hdm2), which in turn decreased the expression of the tumor-suppressor, p53, thereby augmenting transformation that may lead to leukemogenesis.…

    • 820 Words
    • 4 Pages
    Decent Essays
  • Decent Essays

    The aim of gene therapy is to introduce the transgenes or gene into appropriate target cells to restore normal function, therefore, treating and preventing the disease .Similar technologies are being used in gene immunotherapy and DNA vaccines against cancer and other infectious diseases. For a long period of time, current focus with gene therapy is the need for high level of gene expression and to minimize safety issues such as immune reaction against transgene product or transfer vectors and associated cancer…

    • 994 Words
    • 4 Pages
    Decent Essays
  • Decent Essays

    Wnt pathway signaling deregulation contributes to cancer formation and metastasis. Stem cells are likely responsible for the regeneration of breast cancer. There is a linkage between the expression of Wnt1 in human mammary epithelial cells and cancer as it increases stem cell renewal, resistance to apoptosis and failure to senesce. The Wnt pathway is not found in normal stem-like cells. This section is the primary focus of my paper and is a target principle in my focus question: signaling pathways specifically the Wnt-beta catenin pathway.…

    • 1133 Words
    • 5 Pages
    Decent Essays