3. As you are working in the lab, you discover a new TLR (TLR23), which recognizes a pathogen of significant human health concern. Your task, whether or not you choose to accept it, is to fully characterize the molecule. If you are successful, you will definitely finish graduate school and move forward in your career as a scientist. (30 pts)
a. Design an experiment to determine if “TLR23” recognizes a bacteria or a virus. What results do you anticipate and what experimental difficulties will you potentially encounter? TLR’s are toll-like receptors that are associated with resistance to viral, bacterial, parasitic and fungal infections. Flow cytometry would be very useful in being able to fully characterize and analyze the TLR23 molecule. …show more content…
The TLR also has a pathogen recognition domain as well that is made up of LRRs or Leucine Rich Repeats. They are extracellular and made up of twenty to twenty-nine amino acids, majorly leucine. The LRR is also horseshoe shaped to be able to hook onto the pathogen. The TLR23 complex would be located on the macrophage surface and the general platform to start intracellular signaling MyD88, also known as Myeloid Differentiation Factor 88, binds to receptor leading to phosphorylation and activation of IKK. IKK phosphorylates IkB leading to its degradation and the release of NFkB which enter the nucleus. NFkB is a transcription factor that is normally found in cytoplasm. IkB activates transcription of genes for inflammatory cytokines, which are synthesized in the cytoplasm and secreted through the endoplasmic …show more content…
However, different TLRs have signaling domains located extracellularly as well as intracellularly so that they can sense infections inside and outside of the cell. Immunofluorescence would be a great method to be able to see what what specific antibodies have been chemically bound to fluorescent dye within the microtubule cytoskeleton and nuclei. Immunohistochemistry could also be used to selectively take images of antigens and proteins in tissue cells because antibodies bind specifically to antigens in biological tissues through CD3 T-cells. Using immunohistochemistry would make it easy to detect where on the cell surface TLR23 is introduced to the pathogen. The anticipated results would be that when observing the fluorescence of the TLRs that they are found mostly within and around the cell
a. Design an experiment to determine if “TLR23” recognizes a bacteria or a virus. What results do you anticipate and what experimental difficulties will you potentially encounter? TLR’s are toll-like receptors that are associated with resistance to viral, bacterial, parasitic and fungal infections. Flow cytometry would be very useful in being able to fully characterize and analyze the TLR23 molecule. …show more content…
The TLR also has a pathogen recognition domain as well that is made up of LRRs or Leucine Rich Repeats. They are extracellular and made up of twenty to twenty-nine amino acids, majorly leucine. The LRR is also horseshoe shaped to be able to hook onto the pathogen. The TLR23 complex would be located on the macrophage surface and the general platform to start intracellular signaling MyD88, also known as Myeloid Differentiation Factor 88, binds to receptor leading to phosphorylation and activation of IKK. IKK phosphorylates IkB leading to its degradation and the release of NFkB which enter the nucleus. NFkB is a transcription factor that is normally found in cytoplasm. IkB activates transcription of genes for inflammatory cytokines, which are synthesized in the cytoplasm and secreted through the endoplasmic …show more content…
However, different TLRs have signaling domains located extracellularly as well as intracellularly so that they can sense infections inside and outside of the cell. Immunofluorescence would be a great method to be able to see what what specific antibodies have been chemically bound to fluorescent dye within the microtubule cytoskeleton and nuclei. Immunohistochemistry could also be used to selectively take images of antigens and proteins in tissue cells because antibodies bind specifically to antigens in biological tissues through CD3 T-cells. Using immunohistochemistry would make it easy to detect where on the cell surface TLR23 is introduced to the pathogen. The anticipated results would be that when observing the fluorescence of the TLRs that they are found mostly within and around the cell