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PMID: 2745554 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

SPARC, a secreted protein associated with cellular proliferation, inhibits cell spreading in vitro and exhibits Ca+2-dependent binding to the extracellular matrix.

The Journal of cell biology ·Vol. 109 ·No. 1 ·1989-07-00 ·Pages 341-56

Sage H, Vernon RB, Funk SE, Everitt EA, Angello J

Abstract

SPARC (Secreted Protein Acidic and Rich in Cysteine) is a Ca+2-binding glycoprotein that is differentially associated with morphogenesis, remodeling, cellular migration, and proliferation. We show here that exogenous SPARC, added to cells in culture, was associated with profound changes in cell shape, caused rapid, partial detachment of a confluent monolayer, and inhibited spreading of newly plated cells. Bovine aortic endothelial cells, exposed to 2-40 micrograms SPARC/ml per 2 x 10(6) cells, exhibited a rounded morphology in a dose-dependent manner but remained attached to plastic or collagen-coated surfaces. These round cells synthesized protein, uniformly excluded trypan blue, and grew in aggregates after replating in media without SPARC. SPARC caused rounding of bovine endothelial cells, fibroblasts, and smooth muscle cells; however, the cell lines F9, PYS-2, and 3T3 were not affected. The activity of native SPARC was inhibited by heat denaturation and prior incubation with anti-SPARC IgG. The effect of SPARC on endothelial cells appeared to be independent of the rounding phenomenon produced by the peptide GRGDSP. Immunofluorescence localization of SPARC on endothelial cells showed preferential distribution at the leading edges of membranous extensions. SPARC bound Ca+2 in both amino- and carboxyl-terminal (EF-hand) domains and required this cation for maintenance of native structure. Solid-phase binding assays indicated a preferential affinity of native SPARC for several proteins comprising the extracellular matrix, including types III and V collagen, and thrombospondin. This binding was saturable, Ca+2 dependent, and inhibited by anti-SPARC IgG. Endothelial cells also failed to spread on a substrate of native type III collagen complexed with SPARC. We propose that SPARC is an extracellular modulator of Ca+2 and cation-sensitive proteins or proteinases, which facilitates changes in cellular shape and disengagement of cells from the extracellular matrix.

MeSH Terms
Animals Calcium/physiology Carrier Proteins/physiology,ultrastructure Cattle Cell Adhesion Cell Division Cell Movement Cells, Cultured Collagen/metabolism Endothelium, Vascular/cytology Extracellular Matrix/physiology In Vitro Techniques Osteonectin Protein Binding
Chemicals
Carrier Proteins Osteonectin Collagen Calcium
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Sage H
Department of Biological Structure, University of Washington, Seattle 98195.
Vernon R B
Funk S E
Everitt E A
Angello J
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.109.1.341
Published
1989-07-00
Pages
341-56
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2115491
Subset
IM
Grants
NHLBI NIH HHS · HL18645 · United States
External Links
PubMed source DOI: 10.1083/jcb.109.1.341 PMC Full Text View journal details
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