2003 Fiscal Year Final Research Report Summary
Basic study for the bioluminescence reaction on insect luciferases
| Project/Area Number |
12045229
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| Research Category |
Grant-in-Aid for Scientific Research on Priority Areas (A)
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| Allocation Type | Single-year Grants |
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| Review Section |
Science and Engineering
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| Research Institution | Shizuoka University
The National Institute for Advanced Industrial Science and Technology |
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Principal Investigator |
OHMIYA Yoshihiro The National Institute for Advanced Industrial Science and Technology, Research Institute for Cell Engineering, group Leader, セルダイナミクス研究グループ, グループ長 (20223951)
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| Project Period (FY) |
2000
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| Keywords | Active site / Bioluminescence / Light color / Luciferase / Luciferin / Mutants |
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| Research Abstract |
The comparison of click beetle and railroadworm luciferases (pH-insensitive) with firefly luciferases (pH-sensitive) showed a set of conserved residues differing between the two groups which could be involved with the bioluminescence spectra pH sensitivity.The substitution C258V in Pyrocoelia miyako (Pml) firefly luciferase resulted ina red-shifted mutant (λmax= 570 nm). In contrast, the substitution V255C in Ragophthalmus ohbai railroad worm luciferase (Rol) had no effect. Substitution of Thr226 in the green light emitting luciferases of Rol and Pyrearinus termitilluminans (Pyt) click beetle luciferases resulted in red-shifts (12 to 35 nm), whereas the substitution T226N in the red light emitting luciferase of Phrixothrix hirtus (PhRE) railroadworm resulted in a 10 nm blue-shift. In PmL the substitution N230S resulted in a typical red mutant (λ= 611nm). The bioluminescence spectrum of all these luciferase mutants did not showed altered pH-sensitivity nor considerably changed half-bandwidth in relation to the wild-type luciferases. Altogether present data suggest that Thr226 is an important residue for keeping active-site core in both groups of beetle luciferases. The mechanism for bioluminescence color determination between pH-sensitive and pH-insensitive luciferases could be different.
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