title: Cell signaling, internalization, and nuclear localization of the angiotensin converting enzyme in smooth muscle and endothelial cells.
abstract: The angiotensin converting enzyme (ACE) catalyzes the extracellular formation of angiotensin II, and degradation of bradykinin, thus regulating blood pressure and renal handling of electrolytes. We have previously shown that exogenously added ACE elicited transcriptional regulation independent of its enzymatic activity. Because transcriptional regulation generates from protein-DNA interactions within the cell nucleus we have investigated the initial cellular response to exogenous ACE and the putative internalization of the enzyme in smooth muscle cells (SMC) and endothelial cells (EC). The following phenomena were observed when ACE was added to cells in culture: 1) it bound to SMC and EC with high affinity (K(d) = 361.5 +/- 60.5 pM) and with a low binding occupancy (B(max) = 335.0 +/- 14.0 molecules/cell); 2) it triggered cellular signaling resulting in late activation of focal adhesion kinase and SHP2; 3) it modulated platelet-derived growth factor receptor-beta signaling; 4) it was endocytosed by SMC and EC; and 5) it transited through the early endosome, partially occupied the late endosome and the lysosome, and was localized to the nuclei. The incorporation of ACE or a fragment of it into the nuclei reached saturation at 120 min, and was preceded by a lag time of 40 min. Internalized ACE was partially cleaved into small fragments. These results revealed that extracellular ACE modulated cell signaling properties, and that SMC and EC have a pathway for delivery of extracellular ACE to the nucleus, most likely involving cell surface receptor(s) and requiring transit through late endosome/lysosome compartments.
authors.affiliation: Alapis Research Laboratories, Boston, Massachusetts 02118, USA. hlucero@bu.edu
authors.forename: Héctor A
authors.initials: HA
authors.lastname: Lucero
authors.forename: Ekaterina
authors.initials: E
authors.lastname: Kintsurashvili
authors.forename: Maria E
authors.initials: ME
authors.lastname: Marketou
authors.forename: Haralambos
authors.initials: H
authors.lastname: Gavras
mesh_terms: D021581:Active Transport, Cell Nucleus / Q000187:drug effects / Q000502:physiology; D000818:Animals; D002460:Cell Line; D002467:Cell Nucleus / Q000201:enzymology*; D011992:Endosomes / Q000201:enzymology; D042783:Endothelial Cells / Q000166:cytology / Q000201:enzymology*; D004789:Enzyme Activation / Q000187:drug effects / Q000502:physiology; D051417:Focal Adhesion Kinase 1 / Q000378:metabolism; D006801:Humans; D008247:Lysosomes / Q000201:enzymology; D009130:Muscle, Smooth / Q000166:cytology / Q000201:enzymology*; D032389:Myocytes, Smooth Muscle / Q000166:cytology / Q000201:enzymology*; D007703:Peptidyl-Dipeptidase A / Q000378:metabolism* / Q000494:pharmacology; D054592:Protein Tyrosine Phosphatase, Non-Receptor Type 11 / Q000378:metabolism*; D051381:Rats; D020797:Receptor, Platelet-Derived Growth Factor beta / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology*; D014158:Transcription, Genetic / Q000187:drug effects / Q000502:physiology
chemical_list: D020797:Receptor, Platelet-Derived Growth Factor beta; D051417:Focal Adhesion Kinase 1; C496050:PTK2 protein, human; C496052:Ptk2 protein, rat; D054592:Protein Tyrosine Phosphatase, Non-Receptor Type 11; D007703:Peptidyl-Dipeptidase A