Skip Navigation

This Article
Right arrow Full Text Freely available
Right arrow FREE Full Text (PDF) Freely available
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 ISI Web of Science
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 Search for citing articles in:
ISI Web of Science (2)
Right arrowRequest Permissions
Google Scholar
Right arrow Articles by Chen, D.-T.
Right arrow Articles by Lin, A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Chen, D.-T.
Right arrow Articles by Lin, A.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Protein Engineering, Vol. 15, No. 12, 997-1003, December 2002
© 2002 Oxford University Press

Domain swapping in ribonuclease T1 allows the acquisition of double-stranded activity

Dow-Tien Chen and Alan Lin1

Institute of Genetics, National Yang-Ming University, Shih-Pai, Taipei, Taiwan

A mutant of ribonuclease T1 (RNase T1), denoted RNase T{alpha}, that is designed to recognize double-stranded ribonucleic acid was created. RNase T{alpha} carries the structure of RNase T1 except for a part of its loop L3 domain, which has been swapped for a corresponding domain from {alpha}-sarcin. The RNase T{alpha} maintains the pleated ß-sheet structure and retains the guanyl-specific ribonuclease activity of the wild-type RNase T1. A steady-state kinetic study on the RNase T{alpha}-catalyzed transesterification of GpU dinucleoside phosphates reveals a slightly reduced Km value of 6.94x10-7 M. When the stranded specificity is examined, RNase T{alpha} catalyzes the hydrolysis of guanine base not only of single-stranded but also, as by design, of double-stranded RNA. The change of stranded specificity suggests the feasibility of using domain swapping to make a substrate-specific ribonuclease. This study suggests that the loop L3 in RNase T1 can be used as a ‘cassette player’ for inserting a functional domain to make ribonuclease of various specificities.


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
 Nucleic Acids ResHome page
C. Dow-Tien, T. Yuan-Jhih, and L. Alan
Creating a ribonuclease T-tat that preferentially recognizes and hydrolyzes HIV-1 TAR RNA in vitro and in vivo
Nucleic Acids Res., February 11, 2008; 36(3): 963 - 969.
[Abstract] [Full Text] [PDF]

Home page
 Protein Sci.Home page
M. F. Garcia-Mayoral, L. Garcia-Ortega, M. P. Lillo, J. Santoro, A. Martinez Del Pozo, J. G. Gavilanes, M. Rico, and M. Bruix
NMR structure of the noncytotoxic {alpha}-sarcin mutant {Delta}(7-22): The importance of the native conformation of peripheral loops for activity
Protein Sci., April 1, 2004; 13(4): 1000 - 1011.
[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.