Document Detail

Cloning and characterization of an active fragment of luciferase from a luminescent marine alga, Pyrocystis lunula.
MedLine Citation:
PMID:  11950098     Owner:  NLM     Status:  MEDLINE    
Two marine dinoflagellates, Lingulodinium polyedrum and Pyrocystis lunula, emit light in a reaction involving the enzymatic oxidation of its tetrapyrrole luciferin by molecular oxygen. The characteristic properties of P. lunula luciferase have not been clarified, whereas L. polyedrum luciferase, which has three active domains, has been characterized. A cloned partial cDNA of the P. lunula luciferase encodes an active fragment corresponding to part of domain 2 and all of domain 3 of L. polyedrum luciferase. The homology of the amino acid sequence between the two luciferases in domain 3 is about 84.3%. A recombinant His-tagged luciferase fragment containing domain 3 (Mr = 46 kDa) catalyzed the light-emitting oxidation of luciferin (lambdamax = 474 nm). This protein was purified by a single affinity-chromatography procedure. The pH-activity profile and the bioluminescence spectrum of the recombinant enzyme having a third domain are almost identical to those of an extract from P. lunula cultured in vitro. The recombinant enzyme is active at pH 8.0, although the recombinant enzyme derived from the second domain of L. polyedrum luciferase is inactive at pH 8.0. Substitution of Glu-201 by histidine in the third domain of P. lunula luciferase showed a decrease of activity above pH 7.0, suggesting that histidine residues could be responsible for pH-sensitivity in dinoflagellate luciferase.
Hisashi Morishita; Sayumi Ohashi; Takashi Oku; Yoshihiro Nakajima; Satoshi Kojima; Masayuki Ryufuku; Hideshi Nakamura; Yoshihiro Ohmiya
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Photochemistry and photobiology     Volume:  75     ISSN:  0031-8655     ISO Abbreviation:  Photochem. Photobiol.     Publication Date:  2002 Mar 
Date Detail:
Created Date:  2002-04-12     Completed Date:  2002-05-02     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0376425     Medline TA:  Photochem Photobiol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  311-5     Citation Subset:  IM    
Faculty of Education, Shizuoka University, Japan.
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MeSH Terms
Amino Acid Sequence
Base Sequence
Chromatography, Affinity
Cloning, Molecular
DNA Primers
DNA, Complementary
Dinoflagellida / enzymology*
Luciferases / chemistry,  genetics*,  isolation & purification,  metabolism
Marine Biology
Molecular Sequence Data
Sequence Homology, Amino Acid
Reg. No./Substance:
0/DNA Primers; 0/DNA, Complementary; EC 1.13.12.-/Luciferases

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