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Sandbox GGC3
From Proteopedia
(Difference between revisions)
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<StructureSection loadfiles='4G36''4G37' size='340' side='right' caption='Luciferin-4-monooxygenase. The wild-type luciferase in the adenylate-forming conformation with DLSA (PDB 4G36) and the cross-linked luciferase in the second catalytic conformation with DLSA (PDB 4G37)' scene=''> | <StructureSection loadfiles='4G36''4G37' size='340' side='right' caption='Luciferin-4-monooxygenase. The wild-type luciferase in the adenylate-forming conformation with DLSA (PDB 4G36) and the cross-linked luciferase in the second catalytic conformation with DLSA (PDB 4G37)' scene=''> | ||
Firefly luciferase, of the common eastern firefly, is responsible for the ability of the firefly to exhibit bioluminescence. The enzyme luciferin-4-monoxygenase, which catalyzes a multistep oxidative decarboxylation of the luciferyl-AMP intermediate (LH<sub>2</sub>-AMP) to produce bioluminescence, is a part of the ANL superfamily named so for the '''a'''cyl-CoA syntheses, the adenylation domains of the modular '''n'''on-ribosomal peptide synthetases (NRPs), and '''l'''uciferase. | Firefly luciferase, of the common eastern firefly, is responsible for the ability of the firefly to exhibit bioluminescence. The enzyme luciferin-4-monoxygenase, which catalyzes a multistep oxidative decarboxylation of the luciferyl-AMP intermediate (LH<sub>2</sub>-AMP) to produce bioluminescence, is a part of the ANL superfamily named so for the '''a'''cyl-CoA syntheses, the adenylation domains of the modular '''n'''on-ribosomal peptide synthetases (NRPs), and '''l'''uciferase. | ||
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== Function == | == Function == | ||
[[Image:Common_Eastern_Firefly.jpg|thumb|left|The Common Eastern Firefly in a hand emitting a yellow hue, showing bioluminescence.]] | [[Image:Common_Eastern_Firefly.jpg|thumb|left|The Common Eastern Firefly in a hand emitting a yellow hue, showing bioluminescence.]] | ||
The ANL enzymes catalyze two-step reactions: the first an adenylating step in which an acyl-AMP intermediate is produced; the second step in which the adenylate then serves as a substrate for the multistep oxidative decarboxylation of the luciferyl-AMP (LH<sub>2</sub>-AMP) intermediate, resulting in bioluminescence. | The ANL enzymes catalyze two-step reactions: the first an adenylating step in which an acyl-AMP intermediate is produced; the second step in which the adenylate then serves as a substrate for the multistep oxidative decarboxylation of the luciferyl-AMP (LH<sub>2</sub>-AMP) intermediate, resulting in bioluminescence. | ||
| - | ANL enzymes follow a domain alternation strategy for the first adenylation reaction, in which the reaction is catalyzed by one conformation and following the formation of the adenylate intermediate and release of pyrophosphate (PPi), the C-terminal domain undergoes a rotational transformation that is necessary for the second partial reaction. The active site of ANL enzymes resides between a 400-500 residue N-terminal domain and a smaller C-terminal domain of ~110-130 amino acids. Ten conserved regions of these proteins have been termed the A1-A10 motifs which play critical roles in either or both partial reactions. Two lysine residues are required for each partial reaction suggestive that luciferase similarly adopts a rotational transformation for complete catalysis. A mutation of Lys529, the A10 lysine, impairs only the initial adenylation reaction whereas mutation of Lys443 in the A8 region disrupts the oxidative reaction. | + | ANL enzymes follow a domain alternation strategy for the first adenylation reaction, in which the reaction is catalyzed by one conformation and following the formation of the adenylate intermediate and release of pyrophosphate (PPi), the C-terminal domain undergoes a rotational transformation that is necessary for the second partial reaction. The active site of ANL enzymes resides between a 400-500 residue N-terminal domain and a smaller C-terminal domain of ~110-130 amino acids<ref name="Sundlov">Sundlov, J. A., Fontaine, D. M., Southworth, T. L., Branchini, B. R., Gulick, A. M. (2012). Crystal Structure of Firefly Luciferase in a Second Catalytic Conformation Supports a Domain Alternation Mechanism. ''Biochemistry 51''(33), 6493-6495. https://doi.org/10.1021/bi300934s</ref><ref name="Branchini">Branchini, B. R., Murtiashaw, M. H., Magyar, R. A., Anderson, S. M. (2000). The Role of Lysine 529, a Conserved Residue of the Acyl-Adenylate-Forming Enzyme Superfamily, in Firefly Luciferase. ''Biochemistry 39''(18), 5433-5440. https://doi.org/10.1021/bi9928804/>. Ten conserved regions of these proteins have been termed the A1-A10 motifs which play critical roles in either or both partial reactions<ref name="Marahiel">Marahiel, M. A., Stachelhaus, T., Mootz, H. D. (1997). Modular Peptide Synthetases Involved in Nonribosmal Peptide Synthesis. ''Chemical Reviews 97''(7), 2651-2674. https://doi.org/10.1021/cr960029e</ref>. Two lysine residues are required for each partial reaction suggestive that luciferase similarly adopts a rotational transformation for complete catalysis. A mutation of Lys529, the A10 lysine, impairs only the initial adenylation reaction<ref name="Sundlov"/><ref name="Branchini"/> whereas mutation of Lys443 in the A8 region disrupts the oxidative reaction<ref name="Sundlov"/>. |
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== Structural highlights == | == Structural highlights == | ||
| - | The conserved catalytic lysine for the adenylation reaction | + | The conserved catalytic lysine for the adenylation reaction <ref name="Branchini"/><ref name="Branchini1999">Branchini, B. R., Magyar, R. A., Murtiashaw, M. H., Anderson, S. M., Helgerson, L. C., & Zimmer, M. (1999). Site-directed mutagenesis of firefly luciferase active site amino acids: a proposed model for bioluminescence color. ''Biochemistry 38''(40), 13223–13230. https://doi.org/10.1021/bi991181o</ref>, Lys529, interacts with the carbonyl oxygen of the adenylate, the O5 atom that bridges the ribose and sulfamate moiety, and the main chain carbonyl of Gly316. The second conformation observations show that the side chain amine of Lys443 adopts a nearly identical position as Lys529, and Gln448 of the C-terminal domain rotates into the binding pocket where it interacts with a sulfamate oxygen<ref name="Sundlov"/>. Altogether (with the inclusion of an ionic interaction between Glu479 and Arg437), these interactions are responsible for the stabilization of the new C-terminal conformation. |
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| + | (going through changes but math is cool :/)The A8 motif harbors the hinge residue at Lys439 and the antiparallel two stranded β-sheet is directed into the active site of the enzyme. The φ/ψ angles of Lys439 change from −73°/−12° in the structure of wild-type luciferase in the adenylate-forming conformation to −69°/158° in the cross- linked structure.this illustrates that a large component of the conformational change occurs with a rotation of the ψ angle of the hinge residue. Additional torsion angle changes are seen in φ angles for Arg437 and Leu441, although the magnitude of the change is not as large as at the hinge residue Lys439<ref name="Sundlov">. | ||
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| - | (going through changes but math is cool :/)The A8 motif harbors the hinge residue at Lys439 and the antiparallel two stranded β-sheet is directed into the active site of the enzyme. The φ/ψ angles of Lys439 change from −73°/−12° in the structure of wild-type luciferase in the adenylate-forming conformation to −69°/158° in the cross- linked structure.this illustrates that a large component of the conformational change occurs with a rotation of the ψ angle of the hinge residue. Additional torsion angle changes are seen in φ angles for Arg437 and Leu441, although the magnitude of the change is not as large as at the hinge residue Lys439. | ||
| - | <Structure load='4G36' size='350' frame='true' align='left' caption='Subject to change because why not' scene='Insert optional scene name here' /> | ||
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</StructureSection> | </StructureSection> | ||
== References == | == References == | ||
| - | <ref | + | <ref name="Sundlov"/> |
| - | <ref | + | <ref name="Branchini"/> |
| - | <ref | + | <ref name="Marahiel"/> |
| - | <ref | + | <ref name="Branchini1999"/> |
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<references/> | <references/> | ||
Revision as of 12:09, 27 April 2021
Firefly Luciferase
Check over subscripts and underlineee and tttaaarrgggetttt to the right plaaccceee please oh and why no titles for the references :(
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References
- ↑ 1.0 1.1 1.2 1.3 1.4 1.5 Sundlov, J. A., Fontaine, D. M., Southworth, T. L., Branchini, B. R., Gulick, A. M. (2012). Crystal Structure of Firefly Luciferase in a Second Catalytic Conformation Supports a Domain Alternation Mechanism. Biochemistry 51(33), 6493-6495. https://doi.org/10.1021/bi300934s
- ↑ 2.0 2.1 2.2 2.3 Branchini, B. R., Murtiashaw, M. H., Magyar, R. A., Anderson, S. M. (2000). The Role of Lysine 529, a Conserved Residue of the Acyl-Adenylate-Forming Enzyme Superfamily, in Firefly Luciferase. Biochemistry 39(18), 5433-5440. https://doi.org/10.1021/bi9928804/>. Ten conserved regions of these proteins have been termed the A1-A10 motifs which play critical roles in either or both partial reactions<ref>Marahiel, M. A., Stachelhaus, T., Mootz, H. D. (1997). Modular Peptide Synthetases Involved in Nonribosmal Peptide Synthesis. ''Chemical Reviews 97''(7), 2651-2674. https://doi.org/10.1021/cr960029e</li> <li id="cite_note-Branchini1999-2">↑ <sup>[[#cite_ref-Branchini1999_2-0|3.0]]</sup> <sup>[[#cite_ref-Branchini1999_2-1|3.1]]</sup> Branchini, B. R., Magyar, R. A., Murtiashaw, M. H., Anderson, S. M., Helgerson, L. C., & Zimmer, M. (1999). Site-directed mutagenesis of firefly luciferase active site amino acids: a proposed model for bioluminescence color. ''Biochemistry 38''(40), 13223–13230. https://doi.org/10.1021/bi991181o</li> <li id="cite_note-Marahiel">[[#cite_ref-Marahiel_0|↑]] <strong class="error">Cite error: Invalid <code><ref></code> tag; no text was provided for refs named <code>Marahiel</code></strong></li></ol></ref>
