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. 1988 Jan;85(2):577–581. doi: 10.1073/pnas.85.2.577

Molecular mapping of signals in the Qa-2 antigen required for attachment of the phosphatidylinositol membrane anchor.

G L Waneck 1, D H Sherman 1, P W Kincade 1, M G Low 1, R A Flavell 1
PMCID: PMC279594  PMID: 3422441

Abstract

Proteins anchored in the membrane by covalent linkage to phosphatidylinositol (PtdIns) can be released by treatment with purified PtdIns-specific phospholipase C (Ptd-Ins-PLC). A recent survey of leukocyte antigens using flow cytometry has shown that staining of certain Qa antigens was diminished after PtdIns-PLC treatment, but staining of structurally related H-2 antigens was not affected. Therefore, in this study, the sensitivity of cell-surface Qa-2, H-2Kb, and H-2Db to hydrolysis by PtdIns-PLC was investigated biochemically by immunoprecipitation of radioiodinated molecules from cell lysates or supernatants. Qa-2, but not H-2Kb, was released from the surface of PtdIns-PLC-treated C57BL/10 mouse spleen cells and recovered in the cell supernatants. Similar analysis of thymoma cells transfected with cloned C57BL/10 genes showed that cell-surface Qa-2 molecules encoded by a Q7b cDNA and the Q7b or Q9b gene were sensitive to hydrolysis by PtdIns-PLC, whereas the H-2Kb and H-2Db gene products were resistant. Using thymoma cells transfected with hybrid genes constructed by exchanging exons between Q7b and H-2Db, the signals for PtdIns modification were localized to a defined region of Qa-2. This region differs from H-2Db most significantly by the presence of a central aspartate residue in the transmembrane segment and in the length of the cytoplasmic portion.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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