Skip Navigation

This Article
Right arrow Full Text (PDF)
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 Shibata, H.
Right arrow Articles by Oda, J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Shibata, H.
Right arrow Articles by Oda, J.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

J. Biochem, 1998, Vol. 123, No. 1 136-141
© 1998 Japanese Biochemical Society

research-article

Calcium Ion-Dependent Reactivation of a Pseudomonas Lipase by Its Specific Modulating Protein, LipB1

Hiroyuki Shibata, Hiroaki Kato and Jun'ichi Oda2

Institute for Chemical Research, Kyoto University Uji, Kyoto 611

2To whom correspondence should be addressed. Tel: -81-774-38-3230, Fax: +81-774-38-3014, E-mail: oda{at}pclsp2.kuicr.kyoto-u.ac.jp

LipB, the lipase activator protein of Pseudomonas aeruginosa TE3285, was overproduced in Escherichia coli, and purified 4.9-fold over the crude extract in the presence of SDS. The purified LipB reactivated the lipase from P. aeruginosa TE3285 denatured with guanidine hydrochloride, and its reactivation did not involve multiple turnover. In this reactivation, a 1: 1 complex between the lipase and LipB was detected in a cross-linking experiment, suggesting that LipB still binds to the lipase after the reactivation. Calcium ion was essential for the complex formation and the reactivation, and addition of EDTA caused inactivation of the reactivated lipase bound to LipB more rapidly than the native lipase. These findings suggest that LipB could affect the calcium binding to the lipase in the reactivation process. LipB was unable to reactivate lipases from other sources except Pseudomonas sp. 109; this lipase has an amino acid sequence which is 98% identical to that of the lipase from P. aeruginosa TE3285. Thus, it may be concluded that LipB specifically recognizes a unique structural element of the lipase.

1This research was partly supported by Grants-in-Aid for Scientific Research from the Ministry of Education, Science, Sports and Culture of Japan. H.S. was the recipient of Research Fellowships of the Japan Society for the Promotion of Science for Young Scientists.


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


This article has been cited by other articles:


Home page
 Protein Eng Des SelHome page
R. Fujii, Y. Nakagawa, J. Hiratake, A. Sogabe, and K. Sakata
Directed evolution of Pseudomonas aeruginosa lipase for improved amide-hydrolyzing activity
Protein Eng. Des. Sel., February 1, 2005; 18(2): 93 - 101.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
J. Yang, K. Kobayashi, Y. Iwasaki, H. Nakano, and T. Yamane
In Vitro Analysis of Roles of a Disulfide Bridge and a Calcium Binding Site in Activation of Pseudomonas sp. Strain KWI-56 Lipase
J. Bacteriol., January 15, 2000; 182(2): 295 - 302.
[Abstract] [Full Text]

Home page
 J. Biol. Chem.Home page
M. El Khattabi, P. Van Gelder, W. Bitter, and J. Tommassen
Role of the Lipase-specific Foldase of Burkholderia glumae as a Steric Chaperone
J. Biol. Chem., August 25, 2000; 275(35): 26885 - 26891.
[Abstract] [Full Text] [PDF]


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.