Recovery of Sea Star Egg Cell Surface Proteins Released at Fertilization

Methods Mol Biol. 2021:2219:151-161. doi: 10.1007/978-1-0716-0974-3_9.

Abstract

To provide a better understanding of the composition of the egg cell membrane, we describe a method in which proteins and peptides that are either naturally released by the egg or cleaved by sperm proteases can be collected, analyzed, and identified. Such molecules are captured and isolated from the surrounding seawater via biotinylation, before being concentrated by an affinity interaction and subsequently analyzed by western blotting and mass spectrometry.

Keywords: Biotinylation; Blotting; Cell surface; Egg activation; Fertilization; Signal transduction.

Publication types

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

MeSH terms

  • Animals
  • Biotinylation
  • Blotting, Western / methods
  • Cell Culture Techniques / methods
  • Cells, Cultured
  • Egg Proteins / isolation & purification
  • Egg Proteins / metabolism*
  • Electrophoresis, Polyacrylamide Gel / methods
  • Female
  • Fertilization
  • Membrane Proteins / isolation & purification
  • Membrane Proteins / metabolism*
  • Ovum / metabolism
  • Starfish / physiology*

Substances

  • Egg Proteins
  • Membrane Proteins