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Distinct Subcellular Localization of Transiently Expressed Types 1 and 2 Iodothyronine Deiodinases as Determined by Immunofluorescence Confocal Microscopy

Address all correspondence and requests for reprints to: Antonio C. Bianco, M.D., Ph.D., Harvard Institutes of Medicine, 77 Avenue Louis Pasteur, Room 550, Brigham & Women’s Hospital, Boston, Massachusetts 02115.

	Abstract

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We compared the subcellular localization of FLAG-epitope tagged Types 1 and 2 deiodinases (D1 and D2) transiently expressed in human embryonic kidney (HEK-293) and mouse neuroblastoma (NB2A) cells. D2 is an integral membrane protein based on resistance to extraction at pH 11 with the NH₂ terminus in the endoplasmic reticulum (ER). Immunofluorescence confocal microscopy using anti-FLAG and anti-GRP78/BiP antibodies showed the FLAG-D1 signal was found in the periphery of the cells and not co-localized with the ER specific marker GRP78/BiP. On the other hand, FLAG-D2 protein was found in the ER co-localized with the GRP78/BiP protein. These differential distribution patterns indicate subcellular sorting of D1 and D2 is determined by intrinsic protein sequence and can explain the ready access of D2-generated T3 to the nucleus.

Received July 11, 2000.

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