Role of the latent transforming growth factor β binding protein 1 in fibrillin-containing microfibrils in bone cells in vitro and in vivo

Sarah L. Dallas, Douglas R. Keene, Scott P. Bruder, Juha Saharinen, Lynn Y. Sakai, Gregory R. Mundy, Lynda Bonewald

Research output: Contribution to journalArticle

111 Citations (Scopus)

Abstract

Latent transforming growth factor β-binding proteins (LTBPs) are extracellular matrix (ECM) proteins that bind latent transforming growth factor β (TGF-β) and influence its availability in bone and other connective tissues. LTBPs have homology with fibrillins and may have related functions as microfibrillar proteins. However, at present little is known about their structural arrangement in the ECM. By using antibodies against purified LTBP1, against a short peptide in LTBP1, and against epitope-tagged LTBP1 constructs, we have shown colocalization of LTBP1 and fibrillin 1 in microfibrillar structures in the ECM of cultured primary osteoblasts. Immunoelectron microscopy confirmed localization of LTBP1 to 10- to 12-nm microfibrils and suggested an ordered aggregation of LTBP1 into these structures. Early colocalization of LTBP1 with fibronectin suggested a role for fibronectin in the initial assembly of LTBP1 into the matrix; however, in more differentiated osteoblast cultures, LTBP1 and fibronectin 1 were found in distinct fibrillar networks. Overexpression of LTBP1 deletion constructs in osteoblast-like cells showed that N-terminal amino acids 67-467 were sufficient for incorporation into fibrillin-containing microfibrils and suggested that LTBP1 can be produced by cells distant from the site of fibril formation. In embryonic long bones in vivo, LTBP1 and fibrillin 1 colocalized at the surface of newly forming osteoid and bone. However, LTBP1-positive fibrils, which did not contain fibrillin 1, were present in cartilage matrix. These studies show that in addition to regulating TGFβ1, LTBP1 may function as a structural component of connective tissue microfibrils. LTBP1 may therefore be a candidate gene for Marfan-related connective tissue disorders in which linkage to fibrillins has been excluded.

Original languageEnglish (US)
Pages (from-to)68-81
Number of pages14
JournalJournal of Bone and Mineral Research
Volume15
Issue number1
StatePublished - 2000
Externally publishedYes

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Microfibrils
Transforming Growth Factors
Osteoblasts
Fibronectins
Connective Tissue
Carrier Proteins
Bone and Bones
Extracellular Matrix
Immunoelectron Microscopy
Extracellular Matrix Proteins
Cartilage
Epitopes
Amino Acids
Peptides
Antibodies
Genes
In Vitro Techniques
Fibrillin-1
Fibrillins

Keywords

  • Extracellular matrix
  • Fibrillins
  • Latent TGFβ-binding proteins
  • Osteoblasts
  • Transforming growth factor β

ASJC Scopus subject areas

  • Surgery

Cite this

Role of the latent transforming growth factor β binding protein 1 in fibrillin-containing microfibrils in bone cells in vitro and in vivo. / Dallas, Sarah L.; Keene, Douglas R.; Bruder, Scott P.; Saharinen, Juha; Sakai, Lynn Y.; Mundy, Gregory R.; Bonewald, Lynda.

In: Journal of Bone and Mineral Research, Vol. 15, No. 1, 2000, p. 68-81.

Research output: Contribution to journalArticle

Dallas, Sarah L. ; Keene, Douglas R. ; Bruder, Scott P. ; Saharinen, Juha ; Sakai, Lynn Y. ; Mundy, Gregory R. ; Bonewald, Lynda. / Role of the latent transforming growth factor β binding protein 1 in fibrillin-containing microfibrils in bone cells in vitro and in vivo. In: Journal of Bone and Mineral Research. 2000 ; Vol. 15, No. 1. pp. 68-81.
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AU - Mundy, Gregory R.

AU - Bonewald, Lynda

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AB - Latent transforming growth factor β-binding proteins (LTBPs) are extracellular matrix (ECM) proteins that bind latent transforming growth factor β (TGF-β) and influence its availability in bone and other connective tissues. LTBPs have homology with fibrillins and may have related functions as microfibrillar proteins. However, at present little is known about their structural arrangement in the ECM. By using antibodies against purified LTBP1, against a short peptide in LTBP1, and against epitope-tagged LTBP1 constructs, we have shown colocalization of LTBP1 and fibrillin 1 in microfibrillar structures in the ECM of cultured primary osteoblasts. Immunoelectron microscopy confirmed localization of LTBP1 to 10- to 12-nm microfibrils and suggested an ordered aggregation of LTBP1 into these structures. Early colocalization of LTBP1 with fibronectin suggested a role for fibronectin in the initial assembly of LTBP1 into the matrix; however, in more differentiated osteoblast cultures, LTBP1 and fibronectin 1 were found in distinct fibrillar networks. Overexpression of LTBP1 deletion constructs in osteoblast-like cells showed that N-terminal amino acids 67-467 were sufficient for incorporation into fibrillin-containing microfibrils and suggested that LTBP1 can be produced by cells distant from the site of fibril formation. In embryonic long bones in vivo, LTBP1 and fibrillin 1 colocalized at the surface of newly forming osteoid and bone. However, LTBP1-positive fibrils, which did not contain fibrillin 1, were present in cartilage matrix. These studies show that in addition to regulating TGFβ1, LTBP1 may function as a structural component of connective tissue microfibrils. LTBP1 may therefore be a candidate gene for Marfan-related connective tissue disorders in which linkage to fibrillins has been excluded.

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