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Using X-ray crystallography to determine the effect of the antibiotic streptomycin on bacterial protein translation

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Abstract:
My goal was to understand the role of streptomycin, an antibiotic from the aminoglycoside class and the original treatment for tuberculosis, in inhibiting bacterial protein synthesis. With the rise of drug-resistant tuberculosis and other antibiotic-resistant bacterial strains, understanding the functional mechanism of streptomycin and similar antibiotics is becoming increasingly relevant. The Jogl Lab had previously determined via X-ray crystallography that streptomycin created a significant distortion of the 30S ribosomal subunit of Thermus thermophilus in complexes with anticodon stem loops (ASL) and messenger RNA. My hypothesis was that streptomycin disrupted cognate ASL recognition and stabilized near-cognate ASL recognition in the entire 70S ribosome as well, leading to codon misreading and inhibition of protein synthesis. To investigate this theory, I created and executed protocols for the crystallization of wild-type T. thermophilus ribosome with streptomycin and a streptomycin-dependent P90W mutant ribosome for later screening through X-ray diffraction. This task involved determining the optimal conditions for crystal growth through screens as well as the interaction will give us information on codon-anticodon interaction so that new antibiotics may be made or discovered that will still be effective in antibiotic-resistant strains of pathogenic bacteria. This project has significance towards future rational drug design of antibiotics.
Notes:
Senior thesis (ScB)--Brown University, 2017
Concentration: Biochemistry
Concentration: Molecular Biology

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Citation

Rangwala, Aziz M., "Using X-ray crystallography to determine the effect of the antibiotic streptomycin on bacterial protein translation" (2017). Molecular Biology, Cell Biology, and Biochemistry Theses and Dissertations, Chemistry Theses and Dissertations. Brown Digital Repository. Brown University Library. https://doi.org/10.7301/Z0BZ64G1

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