The unique domain as the site on Lyn kinase for its constitutive association with the high affinity receptor for IgE

Becky Marie Vonakis, Huaxian Chen, Hana Haleem-Smith, Henry Metzger

Research output: Contribution to journalArticle

Abstract

Aggregation of the high affinity receptor for IgE (FcεRI) leads to the phosphorylation of tyrosines on the and γ, chains of the receptor by the Src family kinase Lyn. We have studied the interaction between Lyn and the FcεRI in vivo using a transfection-based approach. FcεRI were stably transfected into Chinese hamster ovary cells. The small amount of endogenous Src family kinase was sufficient to phosphorylate receptor tyrosines upon extensive aggregation of FcεRI but not after addition of dimers of IgE. Upon stable co-transfection of Lyn kinase into the cells, dimers were now able to stimulate receptor phosphorylation and the response to more extensive aggregation was enhanced. In contrast, co-transfection with catalytically inactive Lyn inhibited the aggregation-induced phosphorylation by the endogenous kinase, and a quantitatively similar inhibition was observed in cells transfected with the SH4-containing unique domain of Lyn. Consistent with the results of others using alternative approaches, our additional studies using a yeast two-hybrid system detected a direct interaction between intact Lyn or its unique domain and the C-terminal cytoplasmic domain of the chain but not with the receptor's other cytoplasmic domains.

Original languageEnglish (US)
Pages (from-to)24072-24080
Number of pages9
JournalJournal of Biological Chemistry
Volume272
Issue number38
DOIs
StatePublished - Sep 19 1997
Externally publishedYes

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IgE Receptors
Phosphorylation
Transfection
Phosphotransferases
Agglomeration
src-Family Kinases
Association reactions
Dimers
Two-Hybrid System Techniques
Cytoplasmic and Nuclear Receptors
Cricetulus
Immunoglobulin E
Tyrosine
Ovary
Hybrid systems
Yeast
Cells

ASJC Scopus subject areas

  • Biochemistry

Cite this

The unique domain as the site on Lyn kinase for its constitutive association with the high affinity receptor for IgE. / Vonakis, Becky Marie; Chen, Huaxian; Haleem-Smith, Hana; Metzger, Henry.

In: Journal of Biological Chemistry, Vol. 272, No. 38, 19.09.1997, p. 24072-24080.

Research output: Contribution to journalArticle

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AU - Haleem-Smith, Hana

AU - Metzger, Henry

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AB - Aggregation of the high affinity receptor for IgE (FcεRI) leads to the phosphorylation of tyrosines on the and γ, chains of the receptor by the Src family kinase Lyn. We have studied the interaction between Lyn and the FcεRI in vivo using a transfection-based approach. FcεRI were stably transfected into Chinese hamster ovary cells. The small amount of endogenous Src family kinase was sufficient to phosphorylate receptor tyrosines upon extensive aggregation of FcεRI but not after addition of dimers of IgE. Upon stable co-transfection of Lyn kinase into the cells, dimers were now able to stimulate receptor phosphorylation and the response to more extensive aggregation was enhanced. In contrast, co-transfection with catalytically inactive Lyn inhibited the aggregation-induced phosphorylation by the endogenous kinase, and a quantitatively similar inhibition was observed in cells transfected with the SH4-containing unique domain of Lyn. Consistent with the results of others using alternative approaches, our additional studies using a yeast two-hybrid system detected a direct interaction between intact Lyn or its unique domain and the C-terminal cytoplasmic domain of the chain but not with the receptor's other cytoplasmic domains.

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