title: Pasireotide and octreotide stimulate distinct patterns of sst2A somatostatin receptor phosphorylation.
abstract: Pasireotide (SOM230) is currently under clinical evaluation as a successor compound to octreotide for the treatment of acromegaly, Cushing's disease, and carcinoid tumors. Whereas octreotide acts primarily via the sst(2A) somatostatin receptor, pasireotide was designed to exhibit octreotide-like sst(2A) activity combined with enhanced binding to other somatostatin receptor subtypes. In the present study, we used phophosite-specific antibodies to examine agonist-induced phosphorylation of the rat sst(2A) receptor. We show that somatostatin and octreotide stimulate the complete phosphorylation of a cluster of four threonine residues within the cytoplasmic (353)TTETQRT(359) motif in a variety of cultured cell lines in vitro as well as in intact animals in vivo. This phosphorylation was mediated by G protein-coupled receptor kinases (GRK) 2 and 3 and followed by rapid cointernalization of the receptor and ss-arrestin into the same endocytic vesicles. In contrast, pasireotide failed to promote substantial phosphorylation and internalization of the rat sst(2A) receptor. In the presence of octreotide or SS-14, SOM230 showed partial agonist behavior, inhibiting phosphorylation, and internalization of sst(2A). Upon overexpression of GRK2 or GRK3, pasireotide stimulated selective phosphorylation of Thr356 and Thr359 but not of Thr353 or Thr354 within the (353)TTETQRT(359) motif. Pasireotide-mediated phosphorylation led to the formation of relatively unstable beta-arrestin-sst(2A) complexes that dissociated at or near the plasma membrane. Thus, octreotide and pasireotide are equally active in inducing classical G protein-dependent signaling via the sst(2A) somatostatin receptor. Yet, we find that they promote strikingly different patterns of sst(2A) receptor phosphorylation and, hence, stimulate functionally distinct pools of beta-arrestin.
authors.affiliation: Department of Pharmacology and Toxicology, Friedrich-Schiller-University, Drackendorfer Strasse 1, D-07747 Jena, Germany.
authors.forename: Florian
authors.initials: F
authors.lastname: Pöll
authors.forename: Diana
authors.initials: D
authors.lastname: Lehmann
authors.forename: Susann
authors.initials: S
authors.lastname: Illing
authors.forename: Mihaela
authors.initials: M
authors.lastname: Ginj
authors.forename: Stefan
authors.initials: S
authors.lastname: Jacobs
authors.forename: Amelie
authors.initials: A
authors.lastname: Lupp
authors.forename: Ralf
authors.initials: R
authors.lastname: Stumm
authors.forename: Stefan
authors.initials: S
authors.lastname: Schulz
mesh_terms: D000818:Animals; D045163:Antibodies, Phospho-Specific / Q000302:isolation & purification; D019390:Arrestins / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D004705:Endocytosis / Q000187:drug effects; D006801:Humans; D008024:Ligands; D008297:Male; D015282:Octreotide / Q000494:pharmacology*; D010179:Pancreas / Q000187:drug effects / Q000473:pathology; D010766:Phosphorylation / Q000187:drug effects; D010902:Pituitary Gland / Q000187:drug effects / Q000473:pathology; D020033:Protein Isoforms / Q000037:antagonists & inhibitors / Q000378:metabolism; D011499:Protein Processing, Post-Translational / Q000187:drug effects* / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar; D017481:Receptors, Somatostatin / Q000819:agonists / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D013004:Somatostatin / Q000819:agonists* / Q000031:analogs & derivatives* / Q000037:antagonists & inhibitors / Q000494:pharmacology; D013912:Threonine / Q000235:genetics / Q000378:metabolism; D051552:beta-Adrenergic Receptor Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D000071557:beta-Arrestins
chemical_list: D045163:Antibodies, Phospho-Specific; D019390:Arrestins; D008024:Ligands; D020033:Protein Isoforms; D017481:Receptors, Somatostatin; D000071557:beta-Arrestins; C106140:somatostatin receptor sst2A; D013912:Threonine; D013004:Somatostatin; C517782:pasireotide; D051552:beta-Adrenergic Receptor Kinases; D015282:Octreotide