Advertisement
JBC

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


     

Originally published In Press as doi:10.1074/jbc.M300526200 on March 7, 2003

J. Biol. Chem., Vol. 278, Issue 20, 17859-17866, May 16, 2003
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
278/20/17859    most recent
M300526200v1
Right arrow Submit a Letter to Editor
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
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 arrowRequest Permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Shand, J. H.
Right arrow Articles by Allan, G. J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Shand, J. H.
Right arrow Articles by Allan, G. J.
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?

Specific Amino Acid Substitutions Determine the Differential Contribution of the N- and C-terminal Domains of Insulin-like Growth Factor (IGF)-binding Protein-5 in Binding IGF-I*

John H. ShandDagger , James BeattieDagger , Hyuk Song§, Kirsten PhillipsDagger , Sharon M. Kelly||, David J. FlintDagger , and Gordon J. AllanDagger **

From the Dagger  Hannah Research Institute, Ayr KA6 5HL, Scotland, United Kingdom, the § Dana-Farber Cancer Institute and the Department of Pathology, Harvard Medical School, Boston, Massachusetts 02115, and the || Institute of Biomedical and Life Sciences, University of Glasgow, Glasgow G12 8QQ, Scotland, United Kingdom

We have previously reported that two highly conserved amino acids in the C-terminal domain of rat insulin-like growth factor-binding protein (IGFBP)-5, Gly203 and Gln209, are involved in binding to insulin-like growth factor (IGF)-1. Here we report that mutagenesis of both amino acids simultaneously (C-Term mutant) results in a cumulative effect and an even greater reduction in IGF-I binding: 30-fold measured by solution phase IGF binding assay and 10-fold by biosensor analysis. We compared these reductions in ligand binding to the effects of specific mutations of five amino acids in the N-terminal domain (N-Term mutant), which had previously been shown by others to cause a very large reduction in IGF-I binding (1). Our results confirm this as the major IGF-binding site. To prove that the mutations in either N- or C-Term were specific for IGF-I binding, we carried out CD spectroscopy and showed that these alterations did not lead to gross conformational changes in protein structure for either mutant. Combining these mutations in both domains (N+C-Term mutant) has a cumulative effect and leads to a 126-fold reduction in IGF-I binding as measured by biosensor. Furthermore, the equivalent mutations in the C terminus of rat IGFBP-2 (C-Term 2) also results in a significant reduction in IGF-I binding, suggesting that the highly conserved Gly and Gln residues have a conserved IGF-I binding function in all six IGFBPs. Finally, although these residues lie within a major heparin-binding site in IGFBP-5 and -3, we also show that the mutations in C-Term have no effect on heparin binding.

* This work was funded by the Scottish Executive Rural Affairs Department.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 the British Council through the Chevening Scholarship scheme.

** To whom correspondence should be addressed. E-mail: allang@hri.sari.ac.uk.

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
 Mol. Endocrinol.Home page
A. Mukherjee, E. M. Wilson, and P. Rotwein
Insulin-Like Growth Factor (IGF) Binding Protein-5 Blocks Skeletal Muscle Differentiation by Inhibiting IGF Actions
Mol. Endocrinol., January 1, 2008; 22(1): 206 - 215.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Endocrinol.Home page
L. S. Laursen, K. Kjaer-Sorensen, M. H. Andersen, and C. Oxvig
Regulation of Insulin-Like Growth Factor (IGF) Bioactivity by Sequential Proteolytic Cleavage of IGF Binding Protein-4 and -5
Mol. Endocrinol., May 1, 2007; 21(5): 1246 - 1257.
[Abstract] [Full Text] [PDF]

Home page
 EndocrinologyHome page
X. Yan, R. C. Baxter, B. Perbal, and S. M. Firth
The Aminoterminal Insulin-Like Growth Factor (IGF) Binding Domain of IGF Binding Protein-3 Cannot Be Functionally Substituted by the Structurally Homologous Domain of CCN3
Endocrinology, November 1, 2006; 147(11): 5268 - 5274.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
T. Sitar, G. M. Popowicz, I. Siwanowicz, R. Huber, and T. A. Holak
Structural basis for the inhibition of insulin-like growth factors by insulin-like growth factor-binding proteins
PNAS, August 29, 2006; 103(35): 13028 - 13033.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
A. M. Sorrell, J. H. Shand, E. Tonner, M. Gamberoni, P. A. Accorsi, J. Beattie, G. J. Allan, and D. J. Flint
Insulin-like Growth Factor-binding Protein-5 Activates Plasminogen by Interaction with Tissue Plasminogen Activator, Independently of Its Ability to Bind to Plasminogen Activator Inhibitor-1, Insulin-like Growth Factor-I, or Heparin
J. Biol. Chem., April 21, 2006; 281(16): 10883 - 10889.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Pharmacol.Home page
M. M. Kibbey, M. J. Jameson, E. M. Eaton, and S. A. Rosenzweig
Insulin-Like Growth Factor Binding Protein-2: Contributions of the C-Terminal Domain to Insulin-Like Growth Factor-1 Binding
Mol. Pharmacol., March 1, 2006; 69(3): 833 - 845.
[Abstract] [Full Text] [PDF]

Home page
 EndocrinologyHome page
G. J. Allan, E. Tonner, M. Szymanowska, J. H. Shand, S. M. Kelly, K. Phillips, R. A. Clegg, I. F. Gow, J. Beattie, and D. J. Flint
Cumulative Mutagenesis of the Basic Residues in the 201-218 Region of Insulin-Like Growth Factor (IGF)-Binding Protein-5 Results in Progressive Loss of Both IGF-I Binding and Inhibition of IGF-I Biological Action
Endocrinology, January 1, 2006; 147(1): 338 - 349.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
E. Hemers, C. Duval, C. McCaig, M. Handley, G. J. Dockray, and A. Varro
Insulin-Like Growth Factor Binding Protein-5 Is a Target of Matrix Metalloproteinase-7: Implications for Epithelial-Mesenchymal Signaling
Cancer Res., August 15, 2005; 65(16): 7363 - 7369.
[Abstract] [Full Text] [PDF]

Home page
 J Mol EndocrinolHome page
J. Beattie, K. Phillips, J. H Shand, M. Szymanowska, D. J Flint, and G. J Allan
Molecular recognition characteristics in the insulin-like growth factor (IGF)-insulin-like growth factor binding protein -3/5 (IGFBP-3/5) heparin axis
J. Mol. Endocrinol., February 1, 2005; 34(1): 163 - 175.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
X. Yan, B. E. Forbes, K. A. McNeil, R. C. Baxter, and S. M. Firth
Role of N- and C-terminal Residues of Insulin-like Growth Factor (IGF)-binding Protein-3 in Regulating IGF Complex Formation and Receptor Activation
J. Biol. Chem., December 17, 2004; 279(51): 53232 - 53240.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Endocrinol.Home page
S. J. Headey, D. W. Keizer, S. Yao, G. Brasier, P. Kantharidis, L. A. Bach, and R. S. Norton
C-Terminal Domain of Insulin-Like Growth Factor (IGF) Binding Protein-6: Structure and Interaction with IGF-II
Mol. Endocrinol., November 1, 2004; 18(11): 2740 - 2750.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
S. Ramirez-Arcos, V. Greco, H. Douglas, D. Tessier, D. Fan, J. Szeto, J. Wang, and J. R. Dillon
Conserved Glycines in the C Terminus of MinC Proteins Are Implicated in Their Functionality as Cell Division Inhibitors
J. Bacteriol., May 1, 2004; 186(9): 2841 - 2855.
[Abstract] [Full Text] [PDF]

Home page
 J. Cell Sci.Home page
L. J. Cobb, D. A. M. Salih, I. Gonzalez, G. Tripathi, E. J. Carter, F. Lovett, C. Holding, and J. M. Pell
Partitioning of IGFBP-5 actions in myogenesis: IGF-independent anti-apoptotic function
J. Cell Sci., May 1, 2004; 117(9): 1737 - 1746.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
Q. Xu, B. Yan, S. Li, and C. Duan
Fibronectin Binds Insulin-like Growth Factor-binding Protein 5 and Abolishes Its Ligand-dependent Action on Cell Migration
J. Biol. Chem., February 6, 2004; 279(6): 4269 - 4277.
[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.
Advertisement
spacer
Advertisement
Advertisement