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The crystal structures of semi-synthetic aequorins.

Toma S, Chong KT, Nakagawa A, Teranishi K, Inouye S, Shimomura O
Protein Sci 14 409-16 (2005)
Related entries: 1uhi, 1uhj, 1uhk

Cited: 24 times
EuropePMC logo PMID: 15632284

Abstract

The photoprotein aequorin emits light by an intramolecular reaction in the presence of a trace amount of Ca(2+). Semi-synthetic aequorins, produced by replacing the coelenterazine moiety in aequorin with the analogues of coelenterazine, show widely different sensitivities to Ca(2+). To understand the structural basis of the Ca(2+)-sensitivity, we determined the crystal structures of four semi-synthetic aequorins (cp-, i-, br- and n-aequorins) at resolutions of 1.6-1.8 A. In general, the protein structures of these semi-synthetic aequorins are almost identical to native aequorin. Of the four EF-hand domains in the molecule, EF-hand II does not bind Ca(2+), and the loop of EF-hand IV is clearly deformed. It is most likely that the binding of Ca(2+) with EF-hands I and III triggers luminescence. Although little difference was found in the overall structures of aequorins investigated, some significant differences were found in the interactions between the substituents of coelenterazine moiety and the amino acid residues in the binding pocket. The coelenterazine moieties in i-, br-, and n-aequorins have bulky 2-substitutions, which can interfere with the conformational changes of protein structure that follow the binding of Ca(2+) to aequorin. In cp-aequorin, the cyclopentylmethyl group that substitutes for the original 8-benzyl group does not interact hydrophobically with the protein part, giving the coelenterazine moiety more conformational freedom to promote the light-emitting reaction. The differences of various semi-synthetic aequorins in Ca(2+)-sensitivity and reaction rate are explained by the capability of the involved groups and structures to undergo conformational changes in response to the Ca(2+)-binding.

Articles - 1uhh mentioned but not cited (2)

  1. The crystal structures of semi-synthetic aequorins. Toma S, Chong KT, Nakagawa A, Teranishi K, Inouye S, Shimomura O. Protein Sci 14 409-416 (2005)
  2. Crystal structure of semisynthetic obelin-v. Larionova MD, Wu L, Eremeeva EV, Natashin PV, Gulnov DV, Nemtseva EV, Liu D, Liu ZJ, Vysotski ES. Protein Sci 31 454-469 (2022)


Reviews citing this publication (3)

  1. Luminescence of imidazo[1,2-a]pyrazin-3(7H)-one compounds. Teranishi K. Bioorg Chem 35 82-111 (2007)
  2. Fluorescent Protein-photoprotein Fusions and Their Applications in Calcium Imaging. Bakayan A, Domingo B, Vaquero CF, Peyriéras N, Llopis J. Photochem Photobiol 93 448-465 (2017)
  3. Retrospective on the development of aequorin and aequorin-based imaging to visualize changes in intracellular free [Ca(2+) ]. Webb SE, Karplus E, Miller AL. Mol Reprod Dev 82 563-586 (2015)

Articles citing this publication (19)

  1. Cloning, expression, purification and characterization of an isotype of clytin, a calcium-binding photoprotein from the luminous hydromedusa Clytia gregarium. Inouye S. J Biochem 143 711-717 (2008)
  2. Calcium dependence of aequorin bioluminescence dissected by random mutagenesis. Tricoire L, Tsuzuki K, Courjean O, Gibelin N, Bourout G, Rossier J, Lambolez B. Proc Natl Acad Sci U S A 103 9500-9505 (2006)
  3. Blue fluorescent protein from the calcium-sensitive photoprotein aequorin: catalytic properties for the oxidation of coelenterazine as an oxygenase. Inouye S, Sasaki S. FEBS Lett 580 1977-1982 (2006)
  4. NMR analysis of the Mg2+-binding properties of aequorin, a Ca2+-binding photoprotein. Ohashi W, Inouye S, Yamazaki T, Hirota H. J Biochem 138 613-620 (2005)
  5. Spectral components of bioluminescence of aequorin and obelin. Belogurova NV, Kudryasheva NS, Alieva RR, Sizykh AG. J Photochem Photobiol B 92 117-122 (2008)
  6. A unique EF-hand motif in mnemiopsin photoprotein from Mnemiopsis leidyi: implication for its low calcium sensitivity. Jafarian V, Sariri R, Hosseinkhani S, Aghamaali MR, Sajedi RH, Taghdir M, Hassannia S. Biochem Biophys Res Commun 413 164-170 (2011)
  7. Semisynthetic photoprotein reporters for tracking fast Ca(2+) transients. Malikova NP, Borgdorff AJ, Vysotski ES. Photochem Photobiol Sci 14 2213-2224 (2015)
  8. Expression and purification of the calcium binding photoprotein mitrocomin using ZZ-domain as a soluble partner in E. coli cells. Inouye S, Sahara Y. Protein Expr Purif 66 52-57 (2009)
  9. Reconstitution of blue fluorescent protein from recombinant apoaequorin and synthetic coelenteramide. Inouye S, Hosoya T. Biochem Biophys Res Commun 386 617-622 (2009)
  10. Comparison of luminescent immunoassays using biotinylated proteins of aequorin, alkaline phosphatase and horseradish peroxidase as reporters. Inouye S, Sato J. Biosci Biotechnol Biochem 72 3310-3313 (2008)
  11. Expression and characterization of EF-hand I loop mutants of aequorin replaced with other loop sequences of Ca2+-binding proteins: an approach to studying the EF-hand motif of proteins. Inouye S, Sahara-Miura Y. J Biochem 160 59-68 (2016)
  12. Imidazole-assisted catalysis of luminescence reaction in blue fluorescent protein from the photoprotein aequorin. Inouye S, Sasaki S. Biochem Biophys Res Commun 354 650-655 (2007)
  13. Protein Structure Insights into the Bilayer Interactions of the Saposin-Like Domain of Solanum tuberosum Aspartic Protease. Bryksa BC, Yada RY. Sci Rep 7 16911 (2017)
  14. A Novel Catalytic Function of Synthetic IgG-Binding Domain (Z Domain) from Staphylococcal Protein A: Light Emission with Coelenterazine. Inouye S, Sahara-Miura Y. Photochem Photobiol 90 137-144 (2014)
  15. Identification of amino acid residues responsible for high initial luminescence intensity in a calcium-binding photoprotein, clytin-II. Inouye S, Sahara-Miura Y. Biochem Biophys Res Commun 469 300-305 (2016)
  16. Stabilisation of recombinant aequorin by polyols: activity, thermostability and limited proteolysis. Zeinoddini M, Khajeh K, Hosseinkhani S, Saeedinia AR, Robatjazi SM. Appl Biochem Biotechnol 170 273-280 (2013)
  17. RedquorinXS Mutants with Enhanced Calcium Sensitivity and Bioluminescence Output Efficiently Report Cellular and Neuronal Network Activities. Bakayan A, Picaud S, Malikova NP, Tricoire L, Lambolez B, Vysotski ES, Peyriéras N. Int J Mol Sci 21 E7846 (2020)
  18. Slow luminescence kinetics of semi-synthetic aequorin: expression, purification and structure determination of cf3-aequorin. Inouye S, Tomabechi Y, Hosoya T, Sekine SI, Shirouzu M. J Biochem 164 247-255 (2018)
  19. Pretreatment with apoaequorin protects hippocampal CA1 neurons from oxygen-glucose deprivation. Detert JA, Adams EL, Lescher JD, Lyons JA, Moyer JR. PLoS One 8 e79002 (2013)


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