Sandbox GGC3

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==Firefly Luciferase==
==Firefly Luciferase==
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is life imaginary :'~(
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losing my mind B~)
<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 (''Photinus pyralis''), 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 after 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 (''Photinus pyralis''), 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 after the '''a'''cyl-CoA syntheses, the adenylation domains of the modular '''n'''on-ribosomal peptide synthetases (NRPs), and '''l'''uciferase.
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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.
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ANL enzymes follow a domain alternation strategy for the first adenylation reaction, in which the reaction is catalyzed by <scene name='75/752266/4g36/2'>one conformation</scene> 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 <scene name='75/752266/4g37/2'>the second partial reaction</scene>. The <scene name='75/752266/Active_site/1'>active site</scene><ref name=“Branchini”>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> 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>. 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 <scene name='75/752266/Lys529/1'>Lys529</scene>, the A10 lysine, impairs only the initial adenylation reaction<ref name="Sundlov"/> whereas mutation of <scene name='75/752266/Lys443/1'>Lys443</scene> in the A8 region disrupts the oxidative reaction<ref name="Sundlov"/>.
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ANL enzymes follow a domain alternation strategy for the first adenylation reaction, in which the reaction is catalyzed by <scene name='75/752266/4g36/2'>one conformation</scene>, 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 <scene name='75/752266/4g37/2'>the second partial reaction</scene>. The <scene name='75/752266/Active_site/1'>active site</scene><ref name=“Branchini”>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> 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>. 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 <scene name='75/752266/Lys529/1'>Lys529</scene>, the A10 lysine, impairs only the initial adenylation reaction<ref name="Sundlov"/> whereas mutation of <scene name='75/752266/Lys443/1'>Lys443</scene> in the A8 region disrupts the oxidative reaction<ref name="Sundlov"/>.
=== Biochemical Mechanism of LH2-AMP Oxidation===
=== Biochemical Mechanism of LH2-AMP Oxidation===

Revision as of 12:59, 28 April 2021

Firefly Luciferase

losing my mind B~)

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)

Drag the structure with the mouse to rotate

References

  1. ↑ 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
  2. ↑ 2.0 2.1 2.2 2.3 2.4 2.5 2.6 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
  3. ↑ 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
  4. ↑ 4.0 4.1 Branchini, B. R., Southworth, T. L., Murtiahsaw, M. H., Wilkinson, S. R., Khattak, N. F., Rosenberg, J. C., & Zimmer, M. (2005). Mutagenesis Evidence that the Partial Reactions of Firefly Bioluminescence are Catalyzed by Different Conformations of the Luciferase C-Terminal Domain. “Biochemistry 44”(5), 1385-1393. https://doi.org/10.1021/bi047903f
  5. ↑ Oba, Y., Ojika, M., Inouye, S. (2003). Firefly luciferase is a bifunctional enzyme: ATP-dependent monoxygenase and a long chain fatty acyl-CoA synthetase. “FEBS Letters 540”(1-3), 251-254. https://doi.org/10.1016/S0014-5793(03)00272-2
  6. ↑ Nakamura, M., Maki, S., Amano, Y., Ohkita, Y., Niwa, K., Hirano, T., Ohmiya, Y., & Niwa, H. (2005). Firefly luciferase exhibits bimodal action depending on the luciferin chirality. “Biochemical and Biophysical Research Communications, 331”(2), 471–475. https://doi.org/10.1016/j.bbrc.2005.03.202
  7. ↑ 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
  8. ↑ Sala-Newby, G. B., & Campbell, A. K. (1991). Engineering a bioluminescent indicator for cyclic AMP-dependent protein kinase. “The Biochemical Journal”, 279 (Pt 3), 727–732. https://doi.org/10.1042/bj2790727
  9. ↑ de Wet, J. R., Wood, K. V., DeLuca, M., Helinski, D. R., & Subramani, S. (1987). Firefly luciferase gene: structure and expression in mammalian cells. Molecular and cellular biology, 7(2), 725–737. https://doi.org/10.1128/mcb.7.2.725
  10. ↑ de Wet, J. R., Wood, K. V., Helinski, D. R., & DeLuca, M. (1985). Cloning of firefly luciferase cDNA and the expression of active luciferase in Escherichia coli. Proceedings of the National Academy of Sciences of the United States of America, 82(23), 7870–7873. https://doi.org/10.1073/pnas.82.23.7870
  11. ↑ Thorne, N., Shen, M., Lea, W. A., Simeonov, A., Lovell, S., Auld, D. S., & Inglese, J. (2012). Firefly luciferase in chemical biology: a compendium of inhibitors, mechanistic evaluation of chemotypes, and suggested use as a reporter. Chemistry & biology, 19(8), 1060–1072. https://doi.org/10.1016/j.chembiol.2012.07.015
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