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==Identification of Unknown Protein 2QRU== | ==Identification of Unknown Protein 2QRU== | ||
<StructureSection load='2qru' size='340' side=' | <StructureSection load='2qru' size='340' side='left' caption='2QRU' scene=''> | ||
The protein 2QRU is believed to be a hydrolase, functioning best in a near neutral pH environment which could potentially be in muscle cells or in neuronal cells. | |||
</StructureSection> | |||
== '''Introduction''' == | == '''Introduction''' == | ||
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== '''Results''' == | == '''Results''' == | ||
[[Image:SPRITE_pic.png | [[Image:SPRITE_pic.png]] | ||
Figure 1: List of hits of 2QRU from SPRITE (10 of 200 entries shown here). The first column tells what hit each row shows and the second column has the source PDB ID that the program uses to identify each protein in the database. The third column gives a description of the protein and the final column shows the RMSD value of that protein when compared to the search protein of 2QRU. | Figure 1: List of hits of 2QRU from SPRITE (10 of 200 entries shown here). The first column tells what hit each row shows and the second column has the source PDB ID that the program uses to identify each protein in the database. The third column gives a description of the protein and the final column shows the RMSD value of that protein when compared to the search protein of 2QRU. | ||
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Figure 24: UV-vis results at pH 8. There was a steady increase in enzyme activity and then it gradually plateaued. | Figure 24: UV-vis results at pH 8. There was a steady increase in enzyme activity and then it gradually plateaued. | ||
[[Image:PA_6.4.png]] | |||
Figure 25: Protein Activity Assay at pH 6.4. Vmax = 2 x 10-8 M/min. | |||
[[Image:PA_8.png]] | |||
Figure 26: Protein Activity Assay at pH 8. Vmax = 7 x 10-7 M/min. | |||
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== '''Discussion''' == | == '''Discussion''' == | ||
Since the unknown enzyme had a recognizable activity with the common hydrolase substrate, p-nitrophenyl acetate, it is likely that 2QRU is indeed a hydrolase. When unknown protein 2QRU was tested in a pH of ~2, the enzyme completely dissociated from solution indicating it can in no way function in that acidic of an environment in the body. Since the protein worked best in the pH around neutral, it is likely that the enzyme functions as a hydrolase in either neuronal or muscle cells. | Since the unknown enzyme had a recognizable activity with the common hydrolase substrate, p-nitrophenyl acetate, it is likely that 2QRU is indeed a hydrolase. When unknown protein 2QRU was tested in a pH of ~2, the enzyme completely dissociated from solution indicating it can in no way function in that acidic of an environment in the body. Since the protein worked best in the pH around neutral, it is likely that the enzyme functions as a hydrolase in either neuronal or muscle cells. The Vmax at a pH 8 (7 x 10-7 M/min) was slightly higher than the Vmax at pH 6.4 (2 x 10-8 M/min), indicating that although this hydrolase can function in both neuronal and muscle cells, it may favor the environment of muscles cells or be more abundant in muscle cells rather than neuronal cells. | ||
=== Accuracy & Precision of Results === | === Accuracy & Precision of Results === | ||
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== '''Conclusion''' == | == '''Conclusion''' == | ||
Overall, the hypothesis that the protein 2QRU is a type of hydrolase was determined through testing with a general hydrolase-recognized substrate p-nitrophenyl acetate. After testing at three different pH environments, the best pH level for this enzyme seemed to be at a more neutral, between 6.4 and | Overall, the hypothesis that the protein 2QRU is a type of hydrolase was determined through testing with a general hydrolase-recognized substrate p-nitrophenyl acetate. After testing at three different pH environments, the best pH level for this enzyme seemed to be at a more neutral, between 6.4 and 8, being the most active at a pH of 8 which correlates to muscle cells. After computational testing, the best matched substrates are suspected to be 4-nitroacetanilide and alanine-p-nitroanilide. Enzyme activity testing has not been completed thus far with these substrates, but that work could indicate what specific substrates would be best matched with the enzyme providing more specifics into the structure and function of 2QRU. | ||
