Assays for the antiangiogenic and neurotrophic serpin pigment epithelium-derived factor

Methods Enzymol. 2011:499:183-204. doi: 10.1016/B978-0-12-386471-0.00010-9.

Abstract

Pigment epithelium-derived factor (PEDF) is a secreted serpin that exhibits a variety of interesting biological activities. The multifunctional PEDF has neurotrophic and antiangiogenic properties, and acts in retinal differentiation, survival, and maintenance. It is also antitumorigenic and antimetastatic, and has stem cell self-renewal properties. It is widely distributed in the human body and exists in abundance in the eye as a soluble extracellular glycoprotein. Its levels are altered in diseases characterized by retinopathies and angiogenesis. Its mechanisms of neuroprotection and angiogenesis are associated with receptor interactions at cell-surface interfaces and changes in protein expression. This serpin lacks demonstrable serine protease inhibitory activity, but has binding affinity to extracellular matrix components and cell-surface receptors. Here we describe purification protocols, methods to quantify PEDF, and determine interactions with specific molecules, as well as neurotrophic and angiogenesis assays for this multifunctional protein.

Publication types

  • Research Support, N.I.H., Intramural

MeSH terms

  • Ammonium Sulfate / chemistry
  • Animals
  • Aqueous Humor / chemistry
  • Cattle
  • Chromatography, Ion Exchange
  • Enzyme-Linked Immunosorbent Assay
  • Eye Proteins / analysis*
  • Eye Proteins / isolation & purification
  • Immunochemistry
  • Nerve Growth Factors / analysis*
  • Nerve Growth Factors / isolation & purification
  • Serpins / analysis*
  • Serpins / isolation & purification
  • Vitreous Body / chemistry

Substances

  • Eye Proteins
  • Nerve Growth Factors
  • Serpins
  • pigment epithelium-derived factor
  • Ammonium Sulfate