Crystal structure of a pair of follistatin‐like and EF‐hand calcium‐binding domains in BM‐40

BM‐40 (also known as SPARC or osteonectin) is an anti‐adhesive secreted glycoprotein involved in tissue remodelling. Apart from an acidic N‐terminal segment, BM‐40 consists of a follistatin‐like (FS) domain and an EF‐hand calcium‐binding (EC) domain. Here we report the crystal structure at 3.1 Å resolution of the FS–EC domain pair of human BM‐40. The two distinct domains interact through a small interface that involves the EF‐hand pair of the EC domain. Residues implicated in cell binding, inhibition of cell spreading and disassembly of focal adhesions cluster on one face of BM‐40, opposite the binding epitope for collagens and the N‐linked carbohydrate. The elongated FS domain is structurally related to serine protease inhibitors of the Kazal family. Notable differences are an insertion into the inhibitory loop in BM‐40 and a protruding N‐terminal β‐hairpin with striking similarities to epidermal growth factor. This hairpin is likely to act as a rigid spacer in proteins containing tandemly repeated FS domains, such as follistatin and agrin, and forms the heparin‐binding site in follistatin.

[1]  R. Timpl,et al.  Limited Cleavage of Extracellular Matrix Protein BM-40 by Matrix Metalloproteinases Increases Its Affinity for Collagens* , 1997, The Journal of Biological Chemistry.

[2]  O. Podhajcer,et al.  Suppression of SPARC expression by antisense RNA abrogates the tumorigenicity of human melanoma cells , 1997, Nature Medicine.

[3]  E. Sage.  Terms of attachment: SPARC and tumorigenesis , 1997, Nature Medicine.

[4]  L. Reichardt,et al.  Testing the Agrin Hypothesis , 1996, Cell.

[5]  I. Campbell,et al.  Solution Structure of a Pair of Calcium-Binding Epidermal Growth Factor-like Domains: Implications for the Marfan Syndrome and Other Genetic Disorders , 1996, Cell.

[6]  P Bork,et al.  Structure and distribution of modules in extracellular proteins , 1996, Quarterly Reviews of Biophysics.

[7]  T. Springer,et al.  Modulation of Endothelial Cell Adhesion by Hevin, an Acidic Protein Associated with High Endothelial Venules (*) , 1996, The Journal of Biological Chemistry.

[8]  R. Timpl,et al.  Structure of a novel extracellular Ca2+-binding module in BM-40 , 1996, Nature Structural Biology.

[9]  R. Kretsinger.  EF-hands reach out , 1996, Nature Structural Biology.

[10]  R. Timpl,et al.  The C-terminal portion of BM-40 (SPARC/osteonectin) is an autonomously folding and crystallisable domain that binds calcium and collagen IV. , 1995, Journal of molecular biology.

[11]  G. Long,et al.  Role of N-Linked Glycosylation in Human Osteonectin , 1995, The Journal of Biological Chemistry.

[12]  P. Handford,et al.  The structure of a Ca2+-binding epidermal growth factor-like domain: Its role in protein-protein interactions , 1995, Cell.

[13]  G J Kleywegt,et al.  Where freedom is given, liberties are taken. , 1995, Structure.

[14]  A. Bradley,et al.  Multiple defects and perinatal death in mice deficient in follistatin , 1995, Nature.

[15]  N. Ling,et al.  The heparin binding site of follistatin is involved in its interaction with activin. , 1995, Biochemical and biophysical research communications.

[16]  E. Sage,et al.  SPARC mediates focal adhesion disassembly in endothelial cells through a follistatin‐like region and the Ca2+‐binding EF‐hand , 1995, Journal of cellular biochemistry.

[17]  M. Folkman,et al.  Inhibition of endothelial cell proliferation by SPARC is mediated through a Ca2+‐binding EF‐hand sequence , 1995, Journal of cellular biochemistry.

[18]  Collaborative Computational,et al.  The CCP4 suite: programs for protein crystallography. , 1994, Acta crystallographica. Section D, Biological crystallography.

[19]  R. Timpl,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). , 1994, Journal of molecular biology.

[20]  M. Iruela-Arispe,et al.  SPARC is a source of copper-binding peptides that stimulate angiogenesis , 1994, The Journal of cell biology.

[21]  D. Melton,et al.  Follistatin, an antagonist of activin, is expressed in the Spemann organizer and displays direct neuralizing activity , 1994, Cell.

[22]  R. Timpl,et al.  Recombinant expression and structural and binding properties of α1(VI) and α2(VI) chains of human collagen type VI , 1994 .

[23]  J. Navaza,et al.  AMoRe: an automated package for molecular replacement , 1994 .

[24]  E. Sage,et al.  The biology of SPARC, a protein that modulates cell‐matrix interactions , 1994, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[25]  E. Sage,et al.  Specific interaction of SPARC with endothelial cells is mediated through a carboxyl-terminal sequence containing a calcium-binding EF hand. , 1993, The Journal of biological chemistry.

[26]  K. Nose,et al.  Cloning from a mouse osteoblastic cell line of a set of transforming-growth-factor-beta 1-regulated genes, one of which seems to encode a follistatin-related polypeptide. , 1993, European journal of biochemistry.

[27]  J. Schwarzbauer,et al.  The Caenorhabditis elegans homologue of the extracellular calcium binding protein SPARC/osteonectin affects nematode body morphology and mobility. , 1993, Molecular biology of the cell.

[28]  P. Bork,et al.  Epidermal growth factor-like modules , 1993 .

[29]  P. Jollès,et al.  Testican, a multidomain testicular proteoglycan resembling modulators of cell social behaviour. , 1993, European journal of biochemistry.

[30]  J. Thornton,et al.  PROCHECK: a program to check the stereochemical quality of protein structures , 1993 .

[31]  L. Patthy,et al.  Functions of agrin and agrin-related proteins , 1993, Trends in Neurosciences.

[32]  J. Findlay,et al.  Follistatins: More than follicle-stimulating hormone suppressing proteins , 1993, Molecular and Cellular Endocrinology.

[33]  E. Sage,et al.  Differential effects of SPARC and cationic SPARC peptides on DNA synthesis by endothelial cells and fibroblasts , 1993, Journal of cellular physiology.

[34]  R. Timpl,et al.  High-affinity and low-affinity calcium binding and stability of the multidomain extracellular 40-kDa basement membrane glycoprotein (BM-40/SPARC/osteonectin). , 1992, European journal of biochemistry.

[35]  R. Huber,et al.  Natural protein proteinase inhibitors and their interaction with proteinases. , 1992, European journal of biochemistry.

[36]  R. Ross,et al.  The extracellular glycoprotein SPARC interacts with platelet-derived growth factor (PDGF)-AB and -BB and inhibits the binding of PDGF to its receptors. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[37]  K. Titani,et al.  Follistatin, an activin-binding protein, associates with heparan sulfate chains of proteoglycans on follicular granulosa cells. , 1991, The Journal of biological chemistry.

[38]  P. Kraulis.  A program to produce both detailed and schematic plots of protein structures , 1991 .

[39]  N. Ling,et al.  Site-specific mutagenesis of human follistatin. , 1991, Biochemical and biophysical research communications.

[40]  L. Bidou,et al.  Transcription of a quail gene expressed in embryonic retinal cells is shut off sharply at hatching. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[41]  R. Scheller,et al.  Structure and expression of a rat agrin , 1991, Neuron.

[42]  E. Sage,et al.  The Ca2(+)-binding glycoprotein SPARC modulates cell cycle progression in bovine aortic endothelial cells. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[43]  J. Zou,et al.  Improved methods for building protein models in electron density maps and the location of errors in these models. , 1991, Acta crystallographica. Section A, Foundations of crystallography.

[44]  J. Gurd,et al.  Molecular cloning of SC1: A putative brain extracellular matrix glycoprotein showing partial similarity to osteonectin/BM40/SPARC , 1990, Neuron.

[45]  K. Mann,et al.  Structure of human osteonectin based upon analysis of cDNA and genomic sequences. , 1989, Biochemistry.

[46]  M. Young,et al.  Osteonectin cDNA sequence reveals potential binding regions for calcium and hydroxyapatite and shows homologies with both a basement membrane protein (SPARC) and a serine proteinase inhibitor (ovomucoid). , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[47]  M. Paulsson,et al.  Calcium binding domains and calcium-induced conformational transition of SPARC/BM-40/osteonectin, an extracellular glycoprotein expressed in mineralized and nonmineralized tissues. , 1987, Biochemistry.

[48]  N. Ling,et al.  Structural characterization of follistatin: a novel follicle-stimulating hormone release-inhibiting polypeptide from the gonad. , 1987, Molecular endocrinology.

[49]  Randy J. Read,et al.  Crystal and molecular structures of the complex of α-chymotrypsin with its inhibitor Turkey ovomucoid third domain at 1.8 Å resolution , 1987 .

[50]  R. Read.  Improved Fourier Coefficients for Maps Using Phases from Partial Structures with Errors , 1986 .

[51]  R. Doolittle.  The multiplicity of domains in proteins. , 1995, Annual review of biochemistry.

[52]  D. Stuart,et al.  The structure of a Ca(2+)-binding epidermal growth factor-like domain: its role in protein-protein interactions. , 1995, Cell.

[53]  G. Kleywegt,et al.  Halloween ... Masks and Bones , 1994 .

[54]  A. Grollman.  Site specific mutagenesis. , 1990, Progress in clinical and biological research.