Identification and characterization of myeloid translocation gene 16b as a novel A kinase anchoring protein in T lymphocytes

Robynn V. Schillace, Sarah F. Andrews, Greg A. Liberty, Michael Davey, Daniel Carr

Research output: Contribution to journalArticle

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Abstract

Increased levels of intracellular cAMP inhibit T cell activation and proliferation. One mechanism is via activation of the cAMP-dependent protein kinase (PKA). PKA is a broad specificity serine/threonine kinase whose fidelity in signaling is maintained through interactions with A kinase anchoring proteins (AKAPs). AKAPs are adaptor/scaffolding molecules that convey spatial and temporal localization to PKA and other signaling molecules. To determine whether T lymphocytes contain AKAPs that could influence the inflammatory response, PBMCs and Jurkat cells were analyzed for the presence of AKAPs. RII overlay and cAMP pull down assays detected at least six AKAPs. Western blot analyses identified four known AKAPs: AKAP79, AKAP95, AKAP149, and WAVE. Screening of a PMA-stimulated Jurkat cell library identified two additional known AKAPs, AKAP220 and AKAPKL, and one novel AKAP, myeloid translocation gene 16 (MTG16b). Mutational analysis identified the RII binding domain in MTG16b as residues 399-420, and coimmunoprecipitation assays provide strong evidence that MTG16b is an AKAP in vivo. Immunofluorescence and confocal microscopy illustrate distinct subcellular locations of AKAP79, AKAP95, and AKAP149 and suggest colocalization of MTG and RII in the Golgi. These experiments represent the first report of AKAPs in T cells and suggest that MTG16b is a novel AKAP that targets PKA to the Golgi of T lymphocytes.

Original languageEnglish (US)
Pages (from-to)1590-1599
Number of pages10
JournalJournal of Immunology
Volume168
Issue number4
StatePublished - Feb 15 2002
Externally publishedYes

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Protein Kinases
T-Lymphocytes
Genes
Jurkat Cells
Protein-Serine-Threonine Kinases
Protein Transport
Cyclic AMP-Dependent Protein Kinases
Fluorescence Microscopy
Confocal Microscopy
Phosphotransferases
Western Blotting
Cell Proliferation

ASJC Scopus subject areas

  • Immunology

Cite this

Identification and characterization of myeloid translocation gene 16b as a novel A kinase anchoring protein in T lymphocytes. / Schillace, Robynn V.; Andrews, Sarah F.; Liberty, Greg A.; Davey, Michael; Carr, Daniel.

In: Journal of Immunology, Vol. 168, No. 4, 15.02.2002, p. 1590-1599.

Research output: Contribution to journalArticle

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