User:Mitchell Long/Sandbox 1: Difference between revisions
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<scene name='User:Mitchell_Long/Sandbox_1/Luciferase_w_out_cofactor/1'>TextToBeDisplayed</scene> | <scene name='User:Mitchell_Long/Sandbox_1/Luciferase_w_out_cofactor/1'>TextToBeDisplayed</scene> | ||
<scene name='User:Mitchell_Long/Sandbox_1/Hetero_fmn_complex_translucent/1'> | <scene name='User:Mitchell_Long/Sandbox_1/Hetero_fmn_complex_translucent/1'>FMN Positioning</scene> | ||
<scene name='User:Mitchell_Long/Sandbox_1/Hetero_fmn_complex/1'> | <scene name='User:Mitchell_Long/Sandbox_1/Hetero_fmn_complex/1'>Luciferase+FMN</scene> | ||
==Structural Motifs== | ==Structural Motifs== | ||
<p>Structure homology-There is a great deal of sequence homology and structural coservation between the α and β subunits. When superimposed over eachother the barrels of the alpha and beta subunits with a deviation of 0.62Å for 42 equivalent α carbons. The region of the beta subunit that contains the 29 residue deletion with respect to the alpha subunit differs notably in arrangement. In the alpha subunit, the α7a helix is straight and extends toward the beta subunit. The region involved with dimerization, helices α and β and the hairpin loop structure are exceptionally similar in superposition. | <p>'''Structure homology'''-There is a great deal of sequence homology and structural coservation between the α and β subunits. When superimposed over eachother the barrels of the alpha and beta subunits with a deviation of 0.62Å for 42 equivalent α carbons. The region of the beta subunit that contains the 29 residue deletion with respect to the alpha subunit differs notably in arrangement. In the alpha subunit, the α7a helix is straight and extends toward the beta subunit. The region involved with dimerization, helices α and β and the hairpin loop structure are exceptionally similar in superposition. | ||
</p> | </p> | ||
<p>Active Site and Alpha Subunit-the <scene name='User:Mitchell_Long/Sandbox_1/Yellow_sheets/1'>flavin binding pocket</scene> of bacterial luciferase is a large open cavity that is accessible to solvent via an opening located at the C-terminal ends of the ǰ strans of the TIM-barrel structure. During the first step of the oxidation reaction, FMNH<sub>2</sub> binds to the flavin binding pocket and the enzyme undergoes a conformational change that blocks water in the surrounding environment from accessing both the excited peroxydihydroflavin intermediate. Next O<sub>2</sub> and a long chain aldehyde bind to the FMNH<sub>2</sub> luciferase complex and a two step oxidatino reaction occurs. | <p>'''Active Site and Alpha Subunit'''-the <scene name='User:Mitchell_Long/Sandbox_1/Yellow_sheets/1'>flavin binding pocket</scene> of bacterial luciferase is a large open cavity that is accessible to solvent via an opening located at the C-terminal ends of the ǰ strans of the TIM-barrel structure. During the first step of the oxidation reaction, FMNH<sub>2</sub> binds to the flavin binding pocket and the enzyme undergoes a conformational change that blocks water in the surrounding environment from accessing both the excited peroxydihydroflavin intermediate. Next O<sub>2</sub> and a long chain aldehyde bind to the FMNH<sub>2</sub> luciferase complex and a two step oxidatino reaction occurs. | ||
.</p> | .</p> | ||
<scene name='User:Mitchell_Long/Sandbox_1/Hetero_translucent/1'>Heterodimer</scene> | <scene name='User:Mitchell_Long/Sandbox_1/Hetero_translucent/1'>Heterodimer</scene> | ||
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<p>The β subunit-The beta subunit is characterized as a necessary but non-catalytic subunit that stabilizes the catalytic ǯ subunit that is responsible for the oxidation reaction. Both subunits share a great deal of homology | <p>'''The β subunit'''-The beta subunit is characterized as a necessary but non-catalytic subunit that stabilizes the catalytic ǯ subunit that is responsible for the oxidation reaction. Both subunits share a great deal of homology | ||
</p> | </p> | ||
<p>Mobile Loop- Phe2772-thr 288 | <p>'''Mobile Loop'''- Phe2772-thr 288 | ||
(β/α)<SUB>8</SUB> Barrel- The tertiary structure of the α and β subunits is very similar. both subunits fold into a single-domain eight-stranded β/α barrel motif. the two subunits assemble around a parallel four-helix bundle centered on a pseudo 2-fold axis that relates the alpha and beta subunits. | (β/α)<SUB>8</SUB> Barrel- The tertiary structure of the α and β subunits is very similar. both subunits fold into a single-domain eight-stranded β/α barrel motif. the two subunits assemble around a parallel four-helix bundle centered on a pseudo 2-fold axis that relates the alpha and beta subunits. | ||
.</p> | .</p> | ||
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==Applications In Biotechnology== | ==Applications In Biotechnology== | ||
Luciferases are most commonly used as reporter genes by transduction or transfection assays. Reporter genes are inserted into an organism with a gene of interest. This is a powerful method of measuring gene expression because it is non-invasive. Genes for luciferase can paired with an inducible operon. When the gene for luciferase and the gene of interest are incorporated into the host genome, they can "turned on" by induction. | Luciferases are most commonly used as reporter genes by transduction or transfection assays. Reporter genes are inserted into an organism with a gene of interest. This is a powerful method of measuring gene expression because it is non-invasive. Genes for luciferase can paired with an inducible operon. When the gene for luciferase and the gene of interest are incorporated into the host genome, they can "turned on" by induction. Once the desired gene is turned on, gene expression can be determined by the intensity of the light produced by transcription of the gene of interest. | ||
==Quorum Sensing== | ==Quorum Sensing== | ||
