The EF-hand is a highly conserved Ca2+-binding motif found in many cytosolic Ca2+-modulated proteins. Here we report the crystal structure at 2.0 Å resolution of the carboxy-terminal domain of human BM-40 (SPARC, osteonectin), an extracellular matrix protein containing an EF-hand pair. The two EF-hands interact canonically but their detailed structures are unusual. In the first EF-hand a one-residue insertion is accommodated by a cis-peptide bond and by substituting a carboxylate by a peptide carbonyl as a Ca2+ ligand. The second EF-hand is stabilized by a disulphide bond. The EF-hand pair interacts tightly with an amphiphilic amino-terminal helix, reminiscent of target peptide binding by calmodulin. The present structure defines a novel protein module occurring in several other extracellular proteins.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$189.00 per year
only $15.75 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
References
Nakayama, S. & Kretsinger, R.H. Evolution of the EF-hand family of proteins. Annu. Rev. Biophys. biomolec. Struct. 23, 473–507 (1994).
Kretsinger, R.H., Tolbert, D., Nakayama, S. & Pearson, W., The EF-hand, homologs and analogs. In Novel calcium binding proteins (ed Heizmann, C.W.) 17–37 (Springer, Berlin, 1991).
Strynadka, N.C.J. & James, M.N.G. Crystal structures of the helix-loop-helix calcium-binding proteins. A. Rev. Biochem. 58, 951–998 (1989).
McPhalen, C.A., Strynadka, N.C.J. & James, M.N.G. Calcium-binding sites in proteins: a structural perspective. Adv. Prot. Chem. 42, 77–144 (1991).
Kretsinger, R.H. & Nockolds, C.E. Carp muscle calcium-binding protein: structure determination and general description. J. biol. Chem. 9, 3313–3326 (1973).
Finn, B.E. & Forsén, S. The evolving model of calmodulin structure, function and activation. Structure 3, 7–11 (1995).
Crivici, A. & Ikura, M. Molecular and structural basis of target recognition by calmodulin. A. Rev. Biophys. biomol. Struct. 24, 85–116 (1995).
Falke, J.J., Drake, S.K., Hazard, A.L. & Peersen, O.B. Molecular tuning of ion binding to calcium signaling proteins. Quart. Rev. Biophys. 27, 219–290 (1994).
Engel, J., Taylor, W., Paulsson, M., Sage, H. & Hogan, B. Calcium-binding domains and calcium-induced conformational transition of SPARC/BM-40/osteonectin, an extracellular glycoprotein expressed in mineralized and nonmineralized tissues. Biochemistry 26, 6958–6965 (1987).
Bolander, M.E., Young, M.F., Fisher, L.W., Yamada, Y. & Termine, J.D. Osteonectin cDNA sequence reveals potential binding regions for calcium and hydroxyapatite and shows homologies with both a basement membrane protein (SPARC) and a serine protease inhibitor (ovomucoid). Proc. natn. Acad. Sci. U.S.A. 85, 2919–2923 (1988).
Pottgieser, J. et al. Changes in calcium and collagen IV binding caused by mutations in the EF hand and other domains of extracellular matrix protein BM-40 (SPARC, osteonectin). J. molec. Biol. 238, 563–574 (1994).
Termine, J.D. et al. Osteonectin, a bone-specific protein linking mineral to collagen. Cell 26, 99–105 (1981).
Mason, I.J., Taylor, A., Williams, J.G., Sage, H. & Hogan, B.L.M. Evidence from molecular cloning that SPARC, a major product of mouse embryo parietal endoderm, is related to an endothelial cell ‘culture shock’ glycoprotein of Mr 43 000. EMBO J. 5, 1465–1472 (1986).
Mann, K., Deutzmann, R., Paulsson, M. & Timpl, R. Solubilization of protein BM-40 from a basement membrane tumour with chelating agents and evidence for its identity with osteonectin and SPARC. FEBS Lett. 218, 167–172 (1987).
Lane, T.F. & Sage, H. The biology of SPARC, a protein that modulates cell-matrix interactions. FASEB J. 8, 163–173 (1994).
Schwarzbauer, J.E. & Spencer, C.S., The Caenorhabditis elegans homologue of the extracellular calcium binding protein SPARC/osteonectin affects nematode body morphology and mobility. Molec. Biol. Cell 4, 941–952 (1993).
Purcell, L., Gruia-Gray, J., Scanga, S. & Ringuette, M. Developmental anomalies of Xenopus embryos following microinjection of SPARC antibodies. J. exp. Zool. 265, 153–164 (1993).
Patthy, L. Modular exchange principles in proteins. Curr. Opin. struct. Biol. 1, 351–361 (1991).
Maurer, P. et al. High-affinity and low-affinity calcium binding and stability of the multidomain extracellular 40-kDa basement membrane glycoprotein (BM-40/SPARC/osteonectin). Eur. J. Biochem. 205, 233–240 (1992).
Maurer, P. et al. The C-terminal portion of BM-40 (SPARC/osteonectin) is an autonomously folding and crystallisable domain which binds calcium and collagen IV. J. molec. Biol. 253, 347–357 (1995).
Sage, E.H., Bassuk, J.A., Yost, J.C., Folkman, M.J. & Lane, T.F. Inhibition of endothelial cell proliferation by SPARC is mediated through a Ca2+-binding EF-hand sequence. J. cell. Biochem. 57, 127–140 (1995).
Mayer, U., Aumailley, M., Mann, K., Timpl, R. & Engel, J. Calcium-dependent binding of basement membrane protein BM-40 (osteonectin, SPARC) to basement membrane collagen type IV. Eur. J. Biochem. 198, 141–150 (1991).
Xie, X. et al. Structure of the regulatory domain of scallop myosin at 2.8 Å resolution. Nature 368, 306–312 (1994).
Flaherty, K.M., Zozulya, S., Stryer, L. & McKay, D.B. Three-dimensional structure of recoverin, a calcium sensor in vision. Cell 75, 709–716 (1993).
Cook, W.J., Ealick, S.E., Babu, Y.S., Cox, J.A. & Vijay-Kumar, S. Three-dimensional structure of a sarcoplasmic calcium-binding protein from Nereis diversicolor. J. biol. Chem. 266, 652–656 (1991).
Szebenyi, D.M., Obendorf, S.K. & Moffat, K. Structure of vitamin D-dependent calcium-binding protein from bovine intestine. Nature 294, 327–333 (1981).
Chattopadhyaya, R., Meador, W.E., Means, A.R. & Quiocho, F.A. Calmodulin structure refined at 1.7 Å. J. molec. Biol. 228, 1177–1192 (1992).
Ikura, M. et al. Solution structure of a calmodulin-target peptide complex by multidimensional NMR. Science 256, 632–638 (1992).
Meador, W.E., Means, A.R. & Quiocho, F.A. Target enzyme recognition by calmodulin: 2.4 Å structure of a calmodulin-peptide complex. Science 257, 1251–1255 (1992).
Zhang, M., Tanaka, T & Ikura, M. Calcium-induced conformational transition revealed by the solution structure of apo calmodulin. Nature struct. Biology 2, 758–767 (1995).
Kuboniwa, H. et al. Solution structure of calcium-free calmodulin. Nature struct. Biology 2, 768–776 (1995).
Finn et al. Calcium-induced structural changes and domain autonomy in calmodulin. Nature struct. Biology 2, 777–783 (1995).
Clezardin, P. et al. Complex formation of human thrombospondin with osteonectin. Eur. J. Biochem. 175, 275–284 (1988).
Griffith, J.P. et al. X-ray structure of calcineurin inhibited by the immunophilin-immunosuppressant FKBP12-FK506 complex. Cell 82, 507–522 (1995).
Doolittle, R.F. The multiplicity of domains in proteins. A. Rev. Biochem. 64, 287–314 (1995).
Guermah, M. et al. Transcription of a quail gene expressed in embryonic retinal cells is shut off sharply at hatching. Proc. natn. Acad. Sci. U.S.A. 88, 4503–4507 (1991).
Johnston, I.G., Paladino, T., Gurd, J.W. & Brown, I.R. Molecular cloning of SC1: a putative brain extracellular matrix glycoprotein showing partial similarity to osteonectin/BM-40/SPARC. Neuron 2, 165–176 (1990).
Alliel, P.M., Perin, J.P., Jolles, P. & Bonnet, F.J., Testican, a multidomain testicular proteoglycan resembling modulators of cell social behaviour. Eur. J. Biochem. 214, 347–350 (1993).
Shibanuma, M., Mashimo, J., Mita, A., Kuroki, T. & Nose, K. Cloning from a mouse osteoblastic cell line of a set of transforming growth factor β1-regulated genes, one of which seems to encode a follistatin-related polypeptide. Eur. J. Biochem. 217, 13–19 (1993).
Shapiro, L. et al. Structural basis of cell-cell adhesion by cadherins. Nature 374, 327–337 (1995).
Rao, Z. et al. The structure of a Ca2+-binding epidermal growth factor-like domain: its role in protein-protein interactions. Cell 82, 131–141 (1995).
Wendel, M., Sommarin, Y., Bergman, T. & Heinegård, D. Isolation, characterization, and primary structure of a calcium-binding 63-kDa bone protein. J. biol. Chem. 270, 6125–6133 (1995).
Leslie, A.G.W. MOSFLM Users Guide (MRC-LMB, Cambridge, 1994).
Collaborative Computing Project No. 4. The CCP4 suite: programs for protein crystallography. Acta crystallogr. D50, 760–763 (1994).
Jones, T.A., Zou, J.-Y., Cowan, S.W. & Kjeldgaard, M. Improved methods for building protein models in electron density maps and the location of errors in these models. Acta crystallogr. A47, 110–119 (1991).
Brünger, A.T. X-PLOR Version 3.1 A System for Crystallography and NMR (Yale Univ. Press, New Haven, CT, 1992).
Carson, M. Ribbon models of macromolecules. J. molec. Graphics 5, 103–106 (1987).
Lankat-Buttgereit, B., Mann, K., Deutzmann, R., Timpl, R. & Krieg, T. Cloning and complete amino acid sequence of human and murine basement membrane protein BM-40 (SPARC, osteonectin). FEBS Lett. 236, 352–356 (1988).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Hohenester, E., Maurer, P., Hohenadl, C. et al. Structure of a novel extracellular Ca2+-binding module in BM-40. Nat Struct Mol Biol 3, 67–73 (1996). https://doi.org/10.1038/nsb0196-67
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nsb0196-67
