Abstract
Insulin regulated aminopeptidase (IRAP) recognises "AT4-receptor" ligands like angiotensin IV (Ang IV) and peptidomimetics like AL-11. The metabolic stability and high affinity of [3H]AL-11 for catalytically active IRAP allowed its detection in Chinese hamster ovary (CHO-K1) cell membranes in the absence of chelators (Demaegdt et al., 2009). Here, we show that, contrary to [3H]Ang IV, [3H]AL-11 displays high affinity and specificity for IRAP in intact CHO-K1 cells as well. After binding to IRAP at the surface, [3H]AL-11 is effectively internalized by an endocytotic process. Unexpectedly, surface binding and internalization of [3H]AL-11 was not affected by pretreating the cells with Ang IV but declined with AL-11. In the latter case surface expression of IRAP even increased. After elimination of simpler explanations, it is proposed that metabolically stable "AT4-receptor" ligands undergo semi-continuous cycling between the cell surface and endosomal compartments. The in vivo efficacy of stable and unstable "AT4-receptor" ligands could therefore differ.
Original language | English |
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Pages (from-to) | 34-44 |
Number of pages | 11 |
Journal | Molecular and Cellular Endocrinology |
Volume | 339 |
Issue number | 1-2 |
Publication status | Published - 2011 |
Keywords
- IRAP
- CHO cells
- internalization
- radioligand binding
- Ang IV
- AL-11