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PMID: 11170396 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Identification of a motif in the carboxyl terminus of CXCR2 that is involved in adaptin 2 binding and receptor internalization.

Biochemistry ·Vol. 40 ·No. 3 ·2001-01-23 ·Pages 791-800

Fan GH, Yang W, Wang XJ, Qian Q, Richmond A

Abstract

Agonist treatment of cells expressing the chemokine receptor, CXCR2, induces receptor phosphorylation and internalization through a dynamin-dependent mechanism. In the present study, we demonstrate that a carboxyl terminus-truncated mutant of CXCR2 (331T), which no longer undergoes agonist-induced phosphorylation, continues to undergo ligand-induced internalization in HEK293 cells. This mutant receptor exhibits reduced association with beta-arrestin 1 but continues to exhibit association with adaptin 2 alpha and beta subunits. Replacing Leu320-321 and/or Ile323-Leu324 with Ala (LL320,321AA, IL323,324AA, and LLIL320,321,323,324AAAA) in wild-type CXCR2 or 331T causes little change in ligand binding and signaling through Ca(2+) mobilization but greatly impairs the agonist-induced receptor sequestration and ligand-mediated chemotaxis. The LL320,321AA, IL323,324AA, and LLIL320,321,323,324AAAA mutants of CXCR2 exhibit normal binding to beta-arrestin 1 but exhibit decreased binding to adaptin 2alpha and beta. These data demonstrate a role for the LLKIL motif in the carboxyl terminus of CXCR2 in receptor internalization and cell chemotaxis and imply a role for adaptin 2 in the endocytosis of CXCR2.

MeSH Terms
Adaptor Protein Complex alpha Subunits Adaptor Proteins, Vesicular Transport Amino Acid Motifs Amino Acid Sequence Animals Arrestins/metabolism,physiology Cell Line Chemotaxis/genetics,physiology Embryo, Mammalian Humans Kidney Ligands Membrane Proteins/metabolism,physiology Molecular Sequence Data Mutagenesis, Site-Directed Peptide Fragments/chemistry,metabolism,physiology Rats Receptors, Interleukin-8B/agonists,biosynthesis,genetics,metabolism Signal Transduction/genetics Transfection Tumor Cells, Cultured beta-Arrestin 1 beta-Arrestins
Chemicals
ARRB1 protein, human Adaptor Protein Complex alpha Subunits Adaptor Proteins, Vesicular Transport Arrb1 protein, rat Arrestins Ligands Membrane Proteins Peptide Fragments Receptors, Interleukin-8B beta-Arrestin 1 beta-Arrestins
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Fan G H
Veterans Affairs Medical Center, Nashville, Tennessee 37212-2637, USA.
Yang W
Wang X J
Qian Q
Richmond A
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40 references, click to expand
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Article Info
Journal
Biochemistry
Abbr.
Biochemistry
ISSN
0006-2960
DOI
10.1021/bi001661b
Published
2001-01-23
Pages
791-800
Language
English
Region
United States
NLM ID
0370623
PMCID
PMC2664867
Subset
IM
Grants
NCI NIH HHS · R01 CA034590-18 · United States
BLRD VA · IK6 BX005225 · United States
NCI NIH HHS · CA34590 · United States
NCI NIH HHS · R01 CA034590-17 · United States
NCI NIH HHS · R01 CA034590 · United States
NCI NIH HHS · P30 CA068485 · United States
NCI NIH HHS · CA68485 · United States
External Links
PubMed source DOI: 10.1021/bi001661b PMC Full Text View journal details
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