Protein Science
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH
QUICK SEARCH:   [advanced]


     

Published online before print May 7, 2004
Protein Science, DOI: 10.1110/ps.03496004
 Copyright © 2004 The Protein Society
This Article
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
ps.03496004v1
13/6/1566    most recent
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 Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Ohshiro, T.
Right arrow Articles by Izumi, Y.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Ohshiro, T.
Right arrow Articles by Izumi, Y.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

Modification of halogen specificity of a vanadium-dependent bromoperoxidase

Takashi Ohshiro1, Jennifer Littlechild2, Esther Garcia-Rodriguez2, Michail N. Isupov2, Yasuaki Iida1, Takushi Kobayashi1 and Yoshikazu Izumi1

1 Department of Biotechnology, Tottori University, Tottori, 680-8552, Japan
2 Exeter Biocatalysis Centre, School of Biological and Chemical Sciences, University of Exeter, Exeter EX4 4QD, UK

(RECEIVED November 3, 2003; FINAL REVISION January 30, 2004; ACCEPTED February 26, 2004)



Abstract

The halide specificity of vanadium-dependent bromoperoxidase (BPO) from the marine algae, Corallina pilulifera, has been changed by a single amino acid substitution. The residue R397 has been substituted by the other 19 amino acids. The mutant enzymes R397W and R397F showed significant chloroperoxidase (CPO) activity as well as BPO activity. These mutant enzymes were purified and their properties were investigated. The maximal velocities of CPO activities of the R397W and R397F enzymes were 31.2 and 39.2 units/mg, and the Km values for Cl– were 780 mM and 670 mM, respectively. Unlike the native enzyme, both mutant enzymes were inhibited by NaN3. In the case of the R397W enzyme, the incorporation rate of vanadate into the active site was low, compared with the R397F and the wild-type enzyme. These results supported the existence of a specific halogen binding site within the catalytic cleft of vanadium haloperoxidases.

Keywords: haloperoxidase; chloroperoxidase; marine algae; site-directed mutagenesis; vanadate

Reprint request to: Jennifer Littlechild, Exeter Biocatalysis Centre, School of Biological and Chemical Sciences, University of Exeter, Exeter EX4 4QD, UK; e-mail: J.A.Littlechild{at}exeter.ac.uk; fax: 44-1392-263434.

Article published online ahead of print. Article and publication date are at http://www.proteinscience.org/cgi/doi/10.1110/ps.03496004.


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


This article has been cited by other articles:


Home page
 J. Biol. Chem.Home page
Z. Hasan, R. Renirie, R. Kerkman, H. J. Ruijssenaars, A. F. Hartog, and R. Wever
Laboratory-evolved Vanadium Chloroperoxidase Exhibits 100-Fold Higher Halogenating Activity at Alkaline pH: CATALYTIC EFFECTS FROM FIRST AND SECOND COORDINATION SPHERE MUTATIONS
J. Biol. Chem., April 7, 2006; 281(14): 9738 - 9744.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH
Copyright © 2004 by The Protein Society.