Objective: We previously demonstrated that, when expressed in COS-7 cells, L-histidine decarboxylase (HDC), which has neither an amino terminal signal sequence nor a hydrophobic membrane anchor, was localized in the endoplasmic reticulum (ER), although its orientation in the membrane remains to be clarified. Methods & Results: Protease digestion and immunofluorescence analyses of the cells, of which plasma membrane was selectively permeabilized, revealed that the amino terminal 50-kDa portion of HDC is hardly accessible to proteases and antibodies added exogenously from the cytosolic side. Green fluorescent protein fused with the carboxyl terminal 20-kDa region of HDC at its carboxyl terminus exhibited the same characteristics as native HDC. Conclusion: These results indicate that HDC is tightly associated with the ER membrane with its carboxyl terminal region exposed on the cytosolic side.
- Endoplasmic reticulum
- Green fluorescence protein and protease
- Histidine decarboxylase
- Membrane orientation
ASJC Scopus subject areas