A lipid raft environment enhances Lyn kinase activity by protecting the active site tyrosine from dephosphorylation.
The plasma membrane contains ordered lipid domains, commonly called lipid rafts, enriched in cholesterol, sphingolipids, and certain signaling proteins. Lipid rafts play a structural role in signal initiation by the high affinity receptor for IgE. Cross-linking of IgE-receptor complexes by antigen causes their coalescence with lipid rafts, where they are phosphorylated by the Src family tyrosine kinase, Lyn. To understand how lipid rafts participate in functional coupling between Lyn and FcepsilonRI, we investigated whether the lipid raft environment influences the specific activity of Lyn. We used differential detergent solubility and sucrose gradient fractionation to isolate Lyn from raft and nonraft regions of the plasma membrane in the presence or absence of tyrosine phosphatase inhibitors. We show that Lyn recovered from lipid rafts has a substantially higher specific activity than Lyn from nonraft environments. Furthermore, this higher specific activity correlates with increased tyrosine phosphorylation at the active site loop of the kinase domain. Based on these results, we propose that lipid rafts exclude a phosphatase that negatively regulates Lyn kinase activity by constitutive dephosphorylation of the kinase domain tyrosine residue of Lyn. In this model, cross-linking of FcepsilonRI promotes its proximity to active Lyn in a lipid raft environment.
Young
Ryan M
RM
Department of Chemistry and Chemical Biology, Cornell University, Ithaca, New York 14853, USA.
Holowka
David
D
Baird
Barbara
B
D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D003902:Detergents; D004789:Enzyme Activation / Q000502:physiology; D007942:Leukemia, Experimental; D021962:Membrane Microdomains / Q000201:enzymology*; D017830:Octoxynol; D010766:Phosphorylation; D051381:Rats; D014407:Tumor Cells, Cultured; D014443:Tyrosine / Q000378:metabolism; D019061:src-Family Kinases / Q000378:metabolism*
D003902:Detergents; D014443:Tyrosine; D017830:Octoxynol; C060902:lyn protein-tyrosine kinase; D019061:src-Family Kinases