C-terminal motif of human neuropeptide Y4receptor determines internalization and arrestin recruitment.
The human neuropeptide Y4receptor is a rhodopsin-like G protein-coupled receptor (GPCR), which contributes to anorexigenic signals. Thus, this receptor is a highly interesting target for metabolic diseases. As GPCR internalization and trafficking affect receptor signaling and vice versa, we aimed to investigate the molecular mechanism of hY4R desensitization and endocytosis. The role of distinct segments of the hY4R carboxyl terminus was investigated by fluorescence microscopy, binding assays, inositol turnover experiments and bioluminescence resonance energy transfer assays to examine the internalization behavior of hY4R and its interaction with arrestin-3. Based on results of C-terminal deletion mutants and substitution of single amino acids, the motif7.78EESEHLPLSTVHTEVSKGS7.96was identified, with glutamate, threonine and serine residues playing key roles, based on site-directed mutagenesis. Thus, we identified the internalization motif for the human neuropeptide Y4receptor, which regulates arrestin-3 recruitment and receptor endocytosis.
Institute of Biochemistry, Faculty of Biosciences, Pharmacy and Psychology, Leipzig University, Brüderstraße 34, D-04103 Leipzig, Germany.
Lizzy
L
Wanka
Institute of Biochemistry, Faculty of Biosciences, Pharmacy and Psychology, Leipzig University, Brüderstraße 34, D-04103 Leipzig, Germany.
Stefanie
S
Babilon
Institute of Biochemistry, Faculty of Biosciences, Pharmacy and Psychology, Leipzig University, Brüderstraße 34, D-04103 Leipzig, Germany.
Kerstin
K
Burkert
Institute of Biochemistry, Faculty of Biosciences, Pharmacy and Psychology, Leipzig University, Brüderstraße 34, D-04103 Leipzig, Germany.
Karin
K
Mörl
Department of Pharmacology, Vanderbilt University, Nashville, TN 37232, United States.
Vsevolod V
VV
Gurevich
Institute of Biochemistry, Faculty of Biosciences, Pharmacy and Psychology, Leipzig University, Brüderstraße 34, D-04103 Leipzig, Germany. Electronic address: abeck-sickinger@uni-leipzig.de.
Annette G
AG
Beck-Sickinger
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D004705:Endocytosis; D057809:HEK293 Cells; D006801:Humans; D050505:Mutant Proteins / Q000378:metabolism; D017476:Receptors, Neuropeptide Y / Q000737:chemistry* / Q000378:metabolism*; D015203:Reproducibility of Results; D016415:Sequence Alignment; D017384:Sequence Deletion; D013329:Structure-Activity Relationship; D000071560:beta-Arrestin 2 / Q000378:metabolism*
D000596:Amino Acids; D050505:Mutant Proteins; D017476:Receptors, Neuropeptide Y; D000071560:beta-Arrestin 2; C109491:neuropeptide Y4 receptor
Arrestin; G protein-coupled receptors; Internalization; Neuropeptide Y receptors