Skip Navigation


Journal of Biochemistry Advance Access originally published online on February 22, 2008
Journal of Biochemistry 2008 143(5):711-717; doi:10.1093/jb/mvn024
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Supplementary Data
Right arrow All Versions of this Article:
143/5/711    most recent
mvn024v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Request Permissions
Google Scholar
Right arrow Articles by Inouye, S.
PubMed
Right arrow PubMed Citation
Right arrow Articles by Inouye, S.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© 2008 The Japanese Biochemical Society.

Cloning, Expression, Purification and Characterization of an Isotype of Clytin, a Calcium-Binding Photoprotein from the Luminous Hydromedusa Clytia gregarium

Satoshi Inouye*

Yokohama Research Center, Chisso Corporation, 5-1 Okawa, Kanazawa, Yokohama 236-8605, Japan

*To whom correspondence should be addressed. Tel: +81 45 786 5518, Fax: +81 –45 –786 5512, E-mail: sinouye{at}chisso.co.jp

Received January 4, 2008; Accepted February 15, 2008


  Abstract

The cDNA for an isotype of clytin, a calcium-binding photoprotein from the luminous jellyfish Clytia gregarium, was identified and named clytin-II. The histidine-tagged apoprotein of clytin-II expressed into the periplasmic space of Escherichia coli cells was isolated by nickel chelate affinity chromatography. Recombinant clytin-II regenerated from apoprotein by incubation with coelenterazine was purified. The yield of purified clytin-II was 13 mg from 2 l of cultured cells with purity >95%. The luminescence properties of clytin-II were characterized by comparison with clytin-I and aequorin, and semi-synthetic clytin-II was also prepared. The initial luminescence intensity of clytin-II triggered by Ca2+ was 4.5 times higher than that of clytin-I and aequorin, but the luminescence capacity was close to clytin-I and aequorin. Thus, clytin-II is a useful protein, showing high sensitivity in the signal-to-noise ratio of luminescence intensity.

Key Words: aequorin, coelenterazine, luminescence spectrum, protein secretion, isotype clytin

Abbreviations: Apoclytin, apoprotein of clytin; clytin-II, isotype of clytin (clytin-I); OmpA, the outer membrane protein A; EDTA, ethylenediaminetetraacetic acid; DTT, dithiothreitol; Imax, maximum intensity; rlu, relative light units; SDS–PAGE, sodium dodecylsulphate–polyacrylamide gel electrophoresis


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?




Disclaimer:
Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.