Microstructure dependent binding of pigment epithelium derived factor (PEDF) to type I collagen fibrils

Meagan Cauble, Phillip Yang, Ulrich Baumann, Jan M. Gebauer, Bradford G. Orr, Le T. Duong, Mark M. Banaszak Holl

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Pigment epithelium derived factor (PEDF) is a multifunctional extracellular protein. In addition to its known anti-angiogenic and neurotrophic roles in collagen rich tissues, PEDF is thought to be involved in collagen fibril assembly due to its sequence specific binding to the collagen fibril and high expression in regions of active bone formation. In order to image the presence of the protein on the fibrils, PEDF was recombinantly made with a strep tag (strep-PEDF) and then gold nanoparticles conjugated to streptavidin (AuNP) were used as a secondary tag. The gold nanoparticles were detected using phase imaging in tapping mode AFM to image where exogenous PEDF bound in rabbit femur. These findings demonstrate that PEDF binds heterogeneously in cortical rabbit femur. Exogenous PEDF binding was concentrated at areas between microstructures with highly aligned collagen fibrils. Binding was not observed on or within the collagen fibrils themselves.

Original languageEnglish
Pages (from-to)132-139
Number of pages8
JournalJournal of Structural Biology
Volume199
Issue number2
DOIs
Publication statusPublished - 1 Aug 2017
Externally publishedYes

Keywords

  • AFM
  • Collagen
  • PEDF

Cite this

Cauble, Meagan ; Yang, Phillip ; Baumann, Ulrich ; Gebauer, Jan M. ; Orr, Bradford G. ; Duong, Le T. ; Banaszak Holl, Mark M. / Microstructure dependent binding of pigment epithelium derived factor (PEDF) to type I collagen fibrils. In: Journal of Structural Biology. 2017 ; Vol. 199, No. 2. pp. 132-139.
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abstract = "Pigment epithelium derived factor (PEDF) is a multifunctional extracellular protein. In addition to its known anti-angiogenic and neurotrophic roles in collagen rich tissues, PEDF is thought to be involved in collagen fibril assembly due to its sequence specific binding to the collagen fibril and high expression in regions of active bone formation. In order to image the presence of the protein on the fibrils, PEDF was recombinantly made with a strep tag (strep-PEDF) and then gold nanoparticles conjugated to streptavidin (AuNP) were used as a secondary tag. The gold nanoparticles were detected using phase imaging in tapping mode AFM to image where exogenous PEDF bound in rabbit femur. These findings demonstrate that PEDF binds heterogeneously in cortical rabbit femur. Exogenous PEDF binding was concentrated at areas between microstructures with highly aligned collagen fibrils. Binding was not observed on or within the collagen fibrils themselves.",
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Microstructure dependent binding of pigment epithelium derived factor (PEDF) to type I collagen fibrils. / Cauble, Meagan; Yang, Phillip; Baumann, Ulrich; Gebauer, Jan M.; Orr, Bradford G.; Duong, Le T.; Banaszak Holl, Mark M.

In: Journal of Structural Biology, Vol. 199, No. 2, 01.08.2017, p. 132-139.

Research output: Contribution to journalArticleResearchpeer-review

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