title: Carboxyl-terminal domains determine internalization and recycling characteristics of bombesin receptor chimeras.
abstract: To investigate the role of the carboxy terminus in the regulation of the bombesin (BN) receptor, we constructed two chimeric receptors with carboxyl termini transferred from either m3 muscarinic cholinergic (m3 ACh) (BMC) or cholecystokinin A (CCKA) (BCC) receptors and expressed them in Chinese hamster ovary cells. Previous studies showed that agonist treatment caused rapid internalization of CCKA but not m3 ACh receptors in these cells. In the current study we conducted separate analyses of ligand and receptor internalization and analyzed receptor recycling. Ligand internalization was assessed using acid washing. BN and CCKA receptors internalized ligand with 80 +/- 3 and 85 +/- 7% in an acid-resistant compartment at equilibrium. Ligand internalization of chimeric receptors generally assumed the properties of the donor receptors. Thus, BCC receptors internalized ligand to a similar extent as wild-type CCKA receptors (75 +/- 3%), whereas, BMC receptors showed reduced ligand internalization (38 +/- 1%). Receptor internalization was more directly assessed by determining agonist-induced loss of surface binding. BN and CCKA receptors were largely internalized (56 +/- 8 and 50 +/- 7%, respectively). BCC receptors were also extensively internalized (82 +/- 3%). In contrast, BMC receptors were minimally internalized (22 +/- 8%). Receptor recycling was assessed as recovery from agonist induced loss of binding. BN, CCKA, and BMC receptors showed rapid recycling. In contrast, BCC receptors did not recycle. These data indicate that carboxyl-terminal structures determine both internalization of ligand-receptor complexes and subsequent receptor recycling.
authors.affiliation: Department of Physiology, University of Michigan, Ann Arbor 48109-0622, USA.
authors.forename: M J
authors.initials: MJ
authors.lastname: Tseng
authors.forename: K
authors.initials: K
authors.lastname: Detjen
authors.forename: V
authors.initials: V
authors.lastname: Struk
authors.forename: C D
authors.initials: CD
authors.lastname: Logsdon
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D006224:Cricetinae; D008969:Molecular Sequence Data; D018004:Receptors, Bombesin / Q000737:chemistry / Q000378:metabolism*; D011949:Receptors, Cholecystokinin / Q000378:metabolism; D011976:Receptors, Muscarinic / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism*; D013329:Structure-Activity Relationship
chemical_list: D018004:Receptors, Bombesin; D011949:Receptors, Cholecystokinin; D011976:Receptors, Muscarinic; D011993:Recombinant Fusion Proteins