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Isolation and characterization of sea urchin egg lipid rafts and their possible function during fertilization
Author(s) -
Belton Robert J.,
Adams Nikki L.,
Foltz Kathy R.
Publication year - 2001
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/mrd.1034
Subject(s) - biology , lipid raft , microbiology and biotechnology , sea urchin , signal transduction , cytoskeleton , human fertilization , tyrosine kinase , biochemistry , cell , anatomy
Abstract Specialized membrane microdomains called rafts are thought to play a role in many types of cell–cell interactions and signaling. We have investigated the possibility that sea urchin eggs contain these specialized membrane microdomains and if they play a role in signal transduction at fertilization. A low density, TX‐100 insoluble membrane fraction, typical of lipid rafts, was isolated by equilibrium gradient centrifugation. This raft fraction contained proteins distinct from cytoskeletal complexes. The fraction was enriched in tyrosine phosphorylated proteins and contained two proteins known to be involved in signaling during egg activation (an egg Src‐type kinase and PLCγ). This fraction was further characterized as a prototypical raft fraction by the release of proteins in response to in vitro treatment of the rafts with the cholesterol binding drug, methyl‐β‐cyclodextrin (MβCD). Furthermore, treatment of eggs with MβCD inhibited fertilization, suggesting that egg lipid rafts play a physiological role in fertilization. Mol. Reprod. Dev. 59:294–305, 2001. © 2001 Wiley‐Liss, Inc.