1999 Fiscal Year Final Research Report Summary
Study on Molecular Mechanism of Sea Firefly Bioluminescence : Elucidation of Acitive Site in the Luciferase
| Project/Area Number |
10680562
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Bioorganic chemistry
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| Research Institution | The University of Electro-communications |
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Principal Investigator |
NIWA Haruki The University of Electro-communications Applied Physics and Chemistry Professor, 電気通信学部, 教授 (20135297)
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| Co-Investigator(Kenkyū-buntansha) |
MAKI Shojiro The University of Electro-communications Applied Physics and Chemistry Research Associate, 電気通信学部, 助手 (20266349)
OHMIYA Yoshihiro Shizuoka University Department of Biochemistry, 教育学部, 助教授 (20223951)
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| Project Period (FY) |
1998 – 1999
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| Keywords | New Zealand / luminous organism / Latia neritoides / luciferin / luciferase / m-RNA / c-DNA / cloning |
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| Research Abstract |
1) In connection with the structure characterization of post-transrationaly modified proteins by means of mass spectrometry, matrix asisted laser desorption ionization (MALDI)-time of flight (TOF) mass spectrometry was applied to differentiation between a pair of glycosidation site isomeric oligosaccharides, and to determine the phosphorylation site of phosphpeptides. The glycosidation site isomers were found to be distinguished from each other on the basis of post source decay (PSD) spectra from spesific precoursor ions. We found also MALDI-TOF/in source decay (ISD) mass spectrometry to be powerful method for determination of the phosphorylation site of phosphopeptides.
2) We developed a MALDI/TOF mass method for small organic molecules using an ioorganic particle matrix suspended in a viscous dispersant such as liquid paraffin.
3) A photoreactive coelenterazine analogue having an azide group was synthesized, and was found to show similar chemi- and bioluminescence properties to those of natural coelenterazine. Photolysis of the synthesized coelenterazine analogue in the presence of diethylamine gave a azepine derivative derived from the generated nitrene species, as the major product. photoreactive coelenterazine analophotoreactive coelenterazine analo.
4) The small marine ostracod crustacean Vargula hilgendorfi luciferase was purified from the gland-ejected luciferin and luciferase solution in seawater by using size-exclusion chromatography. The luferase was found to be a glycoprotein, which has a molecular mass of 61.7 kDa and pI values of 3.9-5.0, and is a sort of heat stable protein, which maintained 80% activity when incubated for 10 hr at 37 ℃. The activities of the enzyme were maintained in the presence of 0.1 M NaCl, KCl, and MgCl_2.
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Research Products
(10 results)
