Functional analysis of the matricellular protein SPARC with novel monoclonal antibodies

Mariya T. Sweetwyne; Rolf A. Brekken; Gail Workman; Amy D. Bradshaw; Juliet Carbon; Anthony W. Siadak; Carrie Murri; E. Helene Sage

doi:10.1369/jhc.3A6153.2004

Functional analysis of the matricellular protein SPARC with novel monoclonal antibodies

Mariya T. Sweetwyne, Rolf A. Brekken, Gail Workman, Amy D. Bradshaw, Juliet Carbon, Anthony W. Siadak, Carrie Murri, E. Helene Sage

Research output: Contribution to journal › Article › peer-review

38 Scopus citations

Abstract

SPARC (osteonectin, BM-40) is a matricellular glycoprotein that is expressed in many embryogenic and adult tissues undergoing remodeling or repair. SPARC modulates cellular interaction with the extracellular matrix (ECM), inhibits cell adhesion and proliferation, and regulates growth factor activity. To explore further the function and activity of this protein in tissue homeostasis, we have developed several monoclonal antibodies (MAbs) that recognize distinct epitopes on SPARC. The MAbs bind to SPARC with high affinity and identify SPARC by ELISA, Western blotting, immunoprecipitation, immunocytochemistry, and/or immunohistochemistry. The MAbs were also characterized in functional assays for potential alteration of SPARC activity. SPARC binds to collagen I and laminin-1 through an epitope defined by MAb 293; this epitope is not involved in the binding of SPARC to collagen III. The other MAbs did not interfere with the binding of SPARC to collagen I or III or laminin-1. Inhibition of the anti-adhesive effect of SPARC on endothelial cells by MAb 236 was also observed. Functional analysis of SPARC in the presence of these novel MAbs now confirms that the activities ascribed to this matricellular protein can be assigned to discrete subdomains.

Original language	English (US)
Pages (from-to)	723-733
Number of pages	11
Journal	Journal of Histochemistry and Cytochemistry
Volume	52
Issue number	6
DOIs	https://doi.org/10.1369/jhc.3A6153.2004
State	Published - Jun 2004

Keywords

Collagen
Extracellular matrix
Matricellular
Monoclonal antibody
SPARC

ASJC Scopus subject areas

Anatomy
Histology

Access to Document

10.1369/jhc.3A6153.2004

Cite this

@article{7dcfd20d00d545e188cd3dda39388025,

title = "Functional analysis of the matricellular protein SPARC with novel monoclonal antibodies",

abstract = "SPARC (osteonectin, BM-40) is a matricellular glycoprotein that is expressed in many embryogenic and adult tissues undergoing remodeling or repair. SPARC modulates cellular interaction with the extracellular matrix (ECM), inhibits cell adhesion and proliferation, and regulates growth factor activity. To explore further the function and activity of this protein in tissue homeostasis, we have developed several monoclonal antibodies (MAbs) that recognize distinct epitopes on SPARC. The MAbs bind to SPARC with high affinity and identify SPARC by ELISA, Western blotting, immunoprecipitation, immunocytochemistry, and/or immunohistochemistry. The MAbs were also characterized in functional assays for potential alteration of SPARC activity. SPARC binds to collagen I and laminin-1 through an epitope defined by MAb 293; this epitope is not involved in the binding of SPARC to collagen III. The other MAbs did not interfere with the binding of SPARC to collagen I or III or laminin-1. Inhibition of the anti-adhesive effect of SPARC on endothelial cells by MAb 236 was also observed. Functional analysis of SPARC in the presence of these novel MAbs now confirms that the activities ascribed to this matricellular protein can be assigned to discrete subdomains.",

keywords = "Collagen, Extracellular matrix, Matricellular, Monoclonal antibody, SPARC",

author = "Sweetwyne, {Mariya T.} and Brekken, {Rolf A.} and Gail Workman and Bradshaw, {Amy D.} and Juliet Carbon and Siadak, {Anthony W.} and Carrie Murri and Sage, {E. Helene}",

year = "2004",

month = jun,

doi = "10.1369/jhc.3A6153.2004",

language = "English (US)",

volume = "52",

pages = "723--733",

journal = "Journal of Histochemistry and Cytochemistry",

issn = "0022-1554",

publisher = "Histochemical Society Inc.",

number = "6",

}

TY - JOUR

T1 - Functional analysis of the matricellular protein SPARC with novel monoclonal antibodies

AU - Sweetwyne, Mariya T.

AU - Brekken, Rolf A.

AU - Workman, Gail

AU - Bradshaw, Amy D.

AU - Carbon, Juliet

AU - Siadak, Anthony W.

AU - Murri, Carrie

AU - Sage, E. Helene

PY - 2004/6

Y1 - 2004/6

N2 - SPARC (osteonectin, BM-40) is a matricellular glycoprotein that is expressed in many embryogenic and adult tissues undergoing remodeling or repair. SPARC modulates cellular interaction with the extracellular matrix (ECM), inhibits cell adhesion and proliferation, and regulates growth factor activity. To explore further the function and activity of this protein in tissue homeostasis, we have developed several monoclonal antibodies (MAbs) that recognize distinct epitopes on SPARC. The MAbs bind to SPARC with high affinity and identify SPARC by ELISA, Western blotting, immunoprecipitation, immunocytochemistry, and/or immunohistochemistry. The MAbs were also characterized in functional assays for potential alteration of SPARC activity. SPARC binds to collagen I and laminin-1 through an epitope defined by MAb 293; this epitope is not involved in the binding of SPARC to collagen III. The other MAbs did not interfere with the binding of SPARC to collagen I or III or laminin-1. Inhibition of the anti-adhesive effect of SPARC on endothelial cells by MAb 236 was also observed. Functional analysis of SPARC in the presence of these novel MAbs now confirms that the activities ascribed to this matricellular protein can be assigned to discrete subdomains.

AB - SPARC (osteonectin, BM-40) is a matricellular glycoprotein that is expressed in many embryogenic and adult tissues undergoing remodeling or repair. SPARC modulates cellular interaction with the extracellular matrix (ECM), inhibits cell adhesion and proliferation, and regulates growth factor activity. To explore further the function and activity of this protein in tissue homeostasis, we have developed several monoclonal antibodies (MAbs) that recognize distinct epitopes on SPARC. The MAbs bind to SPARC with high affinity and identify SPARC by ELISA, Western blotting, immunoprecipitation, immunocytochemistry, and/or immunohistochemistry. The MAbs were also characterized in functional assays for potential alteration of SPARC activity. SPARC binds to collagen I and laminin-1 through an epitope defined by MAb 293; this epitope is not involved in the binding of SPARC to collagen III. The other MAbs did not interfere with the binding of SPARC to collagen I or III or laminin-1. Inhibition of the anti-adhesive effect of SPARC on endothelial cells by MAb 236 was also observed. Functional analysis of SPARC in the presence of these novel MAbs now confirms that the activities ascribed to this matricellular protein can be assigned to discrete subdomains.

KW - Collagen

KW - Extracellular matrix

KW - Matricellular

KW - Monoclonal antibody

KW - SPARC

UR - http://www.scopus.com/inward/record.url?scp=2542554186&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=2542554186&partnerID=8YFLogxK

U2 - 10.1369/jhc.3A6153.2004

DO - 10.1369/jhc.3A6153.2004

M3 - Article

C2 - 15150281

AN - SCOPUS:2542554186

SN - 0022-1554

VL - 52

SP - 723

EP - 733

JO - Journal of Histochemistry and Cytochemistry

JF - Journal of Histochemistry and Cytochemistry

IS - 6

ER -

Functional analysis of the matricellular protein SPARC with novel monoclonal antibodies

Abstract

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this