The Journal of Cell Biology, Vol 111, Issue 6 3065-3076, Copyright © 1990 by The Rockefeller University Press

ARTICLES

Functional mapping of SPARC: peptides from two distinct Ca+(+)-binding sites modulate cell shape

TF Lane and EH Sage
Department of Biological Structure, University of Washington, Seattle 98195.

Using synthetic peptides, we have identified two distinct regions of the glycoprotein SPARC (Secreted Protein Acidic and Rich in Cysteine) (osteonectin/BM-40) that inhibit cell spreading. One of these sites also contributes to the affinity of SPARC for extracellular matrix components. Peptides representing subregions of SPARC were synthesized and antipeptide antibodies were produced. Immunoglobulin fractions of sera recognizing an NH2-terminal peptide (designated 1.1) blocked SPARC- mediated anti-spreading activity. Furthermore, when peptides were added to newly plated endothelial cells or fibroblasts, peptide 1.1 and a peptide corresponding to the COOH terminal EF-hand domain (designated 4.2) inhibited cell spreading in a dose-dependent manner. These peptides exhibited anti-spreading activity at concentrations from 0.1 to 1 mM. The ability of peptides 1.1 and 4.2 to modulate cell shape was augmented by an inhibitor of protein synthesis and was blocked by specific antipeptide immunoglobulins. In addition to blocking cell spreading, peptide 4.2 competed for binding of [125I]SPARC and exhibited differential affinity for extracellular matrix molecules in solid-phase binding assays. The binding of peptide 4.2 to matrix components was Ca+(+)-dependent and displayed specificities similar to those of native SPARC. These studies demonstrate that both anti- spreading activity and affinity for collagens are functions of unique regions within the SPARC amino acid sequence. The finding that two separate regions of the SPARC protein contribute to its anti-spreading activity lead us to propose that multiple regions of the protein act in concert to regulate the interactions of cells with their extracellular matrix.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Facebook    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

• Weaver, M. S., Workman, G., Sage, E. H. (2008). The Copper Binding Domain of SPARC Mediates Cell Survival in Vitro via Interaction with Integrin {beta}1 and Activation of Integrin-linked Kinase. J. Biol. Chem. 283: 22826-22837 [Abstract] [Full Text]  
• Chlenski, A., Liu, S., Baker, L. J., Yang, Q., Tian, Y., Salwen, H. R., Cohn, S. L. (2004). Neuroblastoma Angiogenesis Is Inhibited with a Folded Synthetic Molecule Corresponding to the Epidermal Growth Factor-Like Module of the Follistatin Domain of SPARC. Cancer Res. 64: 7420-7425 [Abstract] [Full Text]  
• Brekken, R. A., Sullivan, M. M., Workman, G., Bradshaw, A. D., Carbon, J., Siadak, A., Murri, C., Framson, P. E., Sage, E. H. (2004). Expression and Characterization of Murine Hevin (SC1), a Member of the SPARC Family of Matricellular Proteins. J. Histochem. Cytochem. 52: 735-748 [Abstract] [Full Text]  
• Sage, E. H., Reed, M., Funk, S. E., Truong, T., Steadele, M., Puolakkainen, P., Maurice, D. H., Bassuk, J. A. (2003). Cleavage of the Matricellular Protein SPARC by Matrix Metalloproteinase 3 Produces Polypeptides That Influence Angiogenesis. J. Biol. Chem. 278: 37849-37857 [Abstract] [Full Text]  
• Nakada, M., Miyamori, H., Yamashita, J., Sato, H. (2003). Testican 2 Abrogates Inhibition of Membrane-type Matrix Metalloproteinases by Other Testican Family Proteins. Cancer Res. 63: 3364-3369 [Abstract] [Full Text]  
• Chlenski, A., Liu, S., Crawford, S. E., Volpert, O. V., DeVries, G. H., Evangelista, A., Yang, Q., Salwen, H. R., Farrer, R., Bray, J., Cohn, S. L. (2002). SPARC Is a Key Schwannian-derived Inhibitor Controlling Neuroblastoma Tumor Angiogenesis. Cancer Res. 62: 7357-7363 [Abstract] [Full Text]  
• Welt, C., Sidis, Y., Keutmann, H., Schneyer, A. (2002). Activins, Inhibins, and Follistatins: From Endocrinology to Signaling. A Paradigm for the New Millennium. Exp. Biol. Med. 227: 724-752 [Abstract] [Full Text]  
• Yan, Q., Clark, J. I., Wight, T. N., Sage, E. H. (2002). Alterations in the lens capsule contribute to cataractogenesis in SPARC-null mice. J. Cell Sci. 115: 2747-2756 [Abstract] [Full Text]  
• Nakada, M., Yamada, A., Takino, T., Miyamori, H., Takahashi, T., Yamashita, J., Sato, H. (2001). Suppression of Membrane-type 1 Matrix Metalloproteinase (MMP)-mediated MMP-2 Activation and Tumor Invasion by Testican 3 and Its Splicing Variant Gene Product, N-Tes. Cancer Res. 61: 8896-8902 [Abstract] [Full Text]  
• Rodríguez, I. R., Moreira, E. F., Bok, D., Kantorow, M. (2000). Osteonectin/SPARC Secreted by RPE and Localized to the Outer Plexiform Layer of the Monkey Retina. IOVS 41: 2438-2444 [Abstract] [Full Text]  
• Thomas, R., True, L. D., Bassuk, J. A., Lange, P. H., Vessella, R. L. (2000). Differential Expression of Osteonectin/SPARC during Human Prostate Cancer Progression. Clin. Cancer Res. 6: 1140-1149 [Abstract] [Full Text]  
• McKinnon, P. J., McLaughlin, S. K., Kapsetaki, M., Margolskee, R. F. (2000). Extracellular Matrix-Associated Protein Sc1 Is Not Essential for Mouse Development. Mol. Cell. Biol. 20: 656-660 [Abstract] [Full Text]  
• Yan, Q., Sage, E. H. (1999). SPARC, a Matricellular Glycoprotein with Important Biological Functions. J. Histochem. Cytochem. 47: 1495-1506 [Full Text]  
• Bradshaw, A. D., Francki, A., Motamed, K., Howe, C., Sage, E. H. (1999). Primary Mesenchymal Cells Isolated from SPARC-null Mice Exhibit Altered Morphology and Rates of Proliferation. Mol. Biol. Cell 10: 1569-1579 [Abstract] [Full Text]  
• Kupprion, C., Motamed, K., Sage, E. H. (1998). SPARC (BM-40, Osteonectin) Inhibits the Mitogenic Effect of Vascular Endothelial Growth Factor on Microvascular Endothelial Cells. J. Biol. Chem. 273: 29635-29640 [Abstract] [Full Text]  
• Delany, A. M., Canalis, E. (1998). Basic fibroblast growth factor destabilizes osteonectin mRNA in osteoblasts. Am. J. Physiol. Cell Physiol. 274: C734-C740 [Abstract] [Full Text]  
• Soderling, J. A., Reed, M. J., Corsa, A., Sage, E. H. (1997). Cloning and Expression of Murine SC1, a Gene Product Homologous to SPARC. J. Histochem. Cytochem. 45: 823-836 [Abstract] [Full Text]  
• Damjanovski, S., Karp, X., Funk, S., Sage, E. H., Ringuette, M. J. (1997). Ectopic Expression of SPARC in Xenopus Embryos Interferes with Tissue Morphogenesis: Identification of a Bioactive Sequence in the C-terminal EF Hand. J. Histochem. Cytochem. 45: 643-656 [Abstract] [Full Text]  
• McKinnon, P J, Kapsetaki, M, Margolskee, R F (1996). The exon structure of the mouse Sc1 gene is very similar to the mouse Sparc gene.. Genome Res 6: 1077-1083 [Abstract]  
• Girard, J.-P., Springer, T. A. (1996). Modulation of Endothelial Cell Adhesion by Hevin, an Acidic Protein Associated with High Endothelial Venules. J. Biol. Chem. 271: 4511-4517 [Abstract] [Full Text]  
• Murphy-Ullrich, J., Pallero, M., Boerth, N, Greenwood, J., Lincoln, T., Cornwell, T. (1996). Cyclic GMP-dependent protein kinase is required for thrombospondin and tenascin mediated focal adhesion disassembly. J. Cell Sci. 109: 2499-2508 [Abstract]  
• Lian, J. B., Stein, G. S. (1992). Concepts of Osteoblast Growth and Differentiation: Basis for Modulation of Bone Cell Development and Tissue Formation. CROBM 3: 269-305 [Abstract] [Full Text]  
• Sidis, Y., Schneyer, A. L., Sluss, P. M., Johnson, L. N., Keutmann, H. T. (2001). Follistatin: Essential Role for the N-terminal Domain in Activin Binding and Neutralization. J. Biol. Chem. 276: 17718-17726 [Abstract] [Full Text]  

Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents