JBC

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Originally published In Press as doi:10.1074/jbc.M206867200 on December 13, 2002

J. Biol. Chem., Vol. 278, Issue 9, 6896-6904, February 28, 2003
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
278/9/6896    most recent
M206867200v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
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 Odunuga, O. O.
Right arrow Articles by Blatch, G. L.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Odunuga, O. O.
Right arrow Articles by Blatch, G. L.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

Tetratricopeptide Repeat Motif-mediated Hsc70-mSTI1 Interaction
MOLECULAR CHARACTERIZATION OF THE CRITICAL CONTACTS FOR SUCCESSFUL BINDING AND SPECIFICITY*

Odutayo O. OdunugaDagger §, Judith A. HornbyDagger , Christiane Bies, Richard Zimmermann, David J. Pugh||, and Gregory L. BlatchDagger **

From the Dagger  Department of Biochemistry, Microbiology and Biotechnology, Rhodes University, Grahamstown 6140, South Africa,  Medizinische Biochemie und Molekularbiologie, Universität des Saarlandes, D-66421 Homburg, Germany, and the || Department of Biotechnology, University of the Western Cape, Bellville 7535, Cape Town, South Africa

Murine stress-inducible protein 1 (mSTI1) is a co-chaperone that is homologous with the human Hsp70/Hsp90-organizing protein (Hop). Guided by Hop structural data and sequence alignment analyses, we have used site-directed mutagenesis, co-precipitation assays, circular dichroism spectroscopy, steady-state fluorescence, and surface plasmon resonance spectroscopy to both qualitatively and quantitatively characterize the contacts necessary for the N-terminal tetratricopeptide repeat domain (TPR1) of mSTI1 to bind to heat shock cognate protein 70 (Hsc70) and to discriminate between Hsc70 and Hsp90. We have shown that substitutions in the first TPR motif of Lys8 or Asn12 did not affect binding of mSTI1 to Hsc70, whereas double substitution of these residues abrogated binding. A substitution in the second TPR motif of Asn43 lowered but did not abrogate binding. Similarly, a deletion in the second TPR motif coupled with a substitution of Lys8 or Asn12 reduced but did not abrogate binding. These results suggest that mSTI1-Hsc70 interaction requires a network of interactions not only between charged residues in the TPR1 domain of mSTI1 and the EEVD motif of Hsc70 but also outside the TPR domain. We propose that the electrostatic interactions in the first TPR motif made by Lys8 or Asn12 define part of the minimum interactions required for successful mSTI1-Hsc70 interaction. Using a truncated derivative of mSTI1 incapable of binding to Hsp90, we substituted residues on TPR1 potentially involved in hydrophobic contacts with Hsc70. The modified protein had reduced binding to Hsc70 but now showed significant binding capacity for Hsp90. In contrast, topologically equivalent substitutions on a truncated derivative of mSTI1 incapable of binding to Hsc70 did not confer Hsc70 specificity on TPR2A. Our results suggest that binding of Hsc70 to TPR1 is more specific than binding of Hsp90 to TPR2A with serious implications for the mechanisms of mSTI1 interactions with Hsc70 and Hsp90 in vivo.

* This work was supported in part by Volkswagen Programme of Partnerships (Germany) Collaborative Grant 1/77 191 (to R. Z. and G. L. B.), Wellcome Trust (United Kingdom) Grants 053027 and 066705 (to G. L. B.), National Research Foundation (South Africa) Grant 2053542 (to G. L. B.), and a Rhodes University Joint Research Committee grant (to G. L. B.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

§ Supported by Third World Deutscher Akademischer Austauschdienst Predoctoral Scholarship A/99/04651.

** To whom correspondence should be addressed. Tel.: 27-46-6038262; Fax: 27-46-6223984; E-mail: g.blatch@ru.ac.za.

Copyright © 2003 by The American Society for Biochemistry and Molecular Biology, Inc.
Add to CiteULike CiteULike   Add to Complore Complore   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
S. Shimamoto, M. Takata, M. Tokuda, F. Oohira, H. Tokumitsu, and R. Kobayashi
Interactions of S100A2 and S100A6 with the Tetratricopeptide Repeat Proteins, Hsp90/Hsp70-organizing Protein and Kinesin Light Chain
J. Biol. Chem., October 17, 2008; 283(42): 28246 - 28258.
[Abstract] [Full Text] [PDF]

Home page
 GeneticsHome page
F. Ran, M. Bali, and C. A. Michels
Hsp90/Hsp70 Chaperone Machine Regulation of the Saccharomyces MAL-Activator As Determined in Vivo Using Noninducible and Constitutive Mutant Alleles
Genetics, May 1, 2008; 179(1): 331 - 343.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
A. J. Bredemeyer, P. E. Carrigan, T. A. Fehniger, D. F. Smith, and T. J. Ley
Hop Cleavage and Function in Granzyme B-induced Apoptosis
J. Biol. Chem., December 1, 2006; 281(48): 37130 - 37141.
[Abstract] [Full Text] [PDF]

Home page
 Protein Sci.Home page
P. E. Carrigan, L. A. Sikkink, D. F. Smith, and M. Ramirez-Alvarado
Domain:domain interactions within Hop, the Hsp70/Hsp90 organizing protein, are required for protein stability and structure
Protein Sci., March 1, 2006; 15(3): 522 - 532.
[Abstract] [Full Text] [PDF]

Home page
 GeneticsHome page
G. Flom, J. Weekes, J. J. Williams, and J. L. Johnson
Effect of Mutation of the Tetratricopeptide Repeat and Asparatate-Proline 2 Domains of Sti1 on Hsp90 Signaling and Interaction in Saccharomyces cerevisiae
Genetics, January 1, 2006; 172(1): 41 - 51.
[Abstract] [Full Text] [PDF]

Home page
 Sci SignalHome page
D. C. Love and J. A. Hanover
The Hexosamine Signaling Pathway: Deciphering the "O-GlcNAc Code"
Sci. Signal., November 29, 2005; 2005(312): re13 - re13.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
H. M. Jones, K. L. Hamilton, and D. C. Devor
Role of an S4-S5 Linker Lysine in the Trafficking of the Ca2+-activated K+ Channels IK1 and SK3
J. Biol. Chem., November 4, 2005; 280(44): 37257 - 37265.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
Y. Song and D. C. Masison
Independent Regulation of Hsp70 and Hsp90 Chaperones by Hsp70/Hsp90-organizing Protein Sti1 (Hop1)
J. Biol. Chem., October 7, 2005; 280(40): 34178 - 34185.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
P. E. Carrigan, D. L. Riggs, M. Chinkers, and D. F. Smith
Functional Comparison of Human and Drosophila Hop Reveals Novel Role in Steroid Receptor Maturation
J. Biol. Chem., March 11, 2005; 280(10): 8906 - 8911.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
P. E. Carrigan, G. M. Nelson, P. J. Roberts, J. Stoffer, D. L. Riggs, and D. F. Smith
Multiple Domains of the Co-chaperone Hop Are Important for Hsp70 Binding
J. Biol. Chem., April 16, 2004; 279(16): 16185 - 16193.
[Abstract] [Full Text] [PDF]

Home page
 Protein Eng Des SelHome page
A. L. Cortajarena, T. Kajander, W. Pan, M. J. Cocco, and L. Regan
Protein design to understand peptide ligand recognition by tetratricopeptide repeat proteins
Protein Eng. Des. Sel., April 1, 2004; 17(4): 399 - 409.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
S. Tobaben, F. Varoqueaux, N. Brose, B. Stahl, and G. Meyer
A Brain-specific Isoform of Small Glutamine-rich Tetratricopeptide Repeat-containing Protein Binds to Hsc70 and the Cysteine String Protein
J. Biol. Chem., October 3, 2003; 278(40): 38376 - 38383.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
 All ASBMB Journals   Molecular and Cellular Proteomics 
 Journal of Lipid Research   ASBMB Today 
Copyright © 2003 by the American Society for Biochemistry and Molecular Biology.