作者
Elaine Huston, Linda Pooley, Pascale Julien, Grant Scotland, Ian McPhee, Michael Sullivan, Graeme Bolger, Miles D Houslay
发表日期
1996/12/6
期刊
Journal of Biological Chemistry
卷号
271
期号
49
页码范围
31334-31344
出版商
Elsevier
简介
Transfection of COS7 cells with a plasmid encoding the human cyclic AMP-specific PDE4A phosphodiesterase PDE-46 (HSPDE4A4B) led to the expression of a rolipram-inhibited PDE4 activity, which contributed ∼96% of the total COS cell PDE activity. A fusion protein was generated which encompassed residues (788-886) at the extreme C terminus of PDE-46 and was used to generate an antiserum that detected PDE-46 in transfected COS7 cells. Immunoblotting studies identified PDE-46 as a ∼125-kDa species that was associated with both the soluble and particulate fractions. The relative Vmax of particulate PDE-46 was ∼56% that of cytosolic PDE-46. Particulate PDE-46 was not solubilized using Triton X-100 or high NaCl concentrations. Immunofluorescence analysis by laser scanning confocal microscopy showed that PDE-46 was located at discrete margins of the cell, indicative of association with …
引用总数
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