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(Received for publication, February 12, 1997)
From the Departments of Biochemistry and Molecular Pharmacology and
Microbiology and Immunology, Kimmel Cancer Institute, Thomas
Jefferson University, Philadelphia, Pennsylvania 19107
We have recently demonstrated that the nonvisual
arrestins,
Volume 272, Number 23,
Issue of June 6, 1997
pp. 15011-15016
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.
LOCALIZATION OF THE CLATHRIN BINDING DOMAIN OF NONVISUAL
ARRESTINS TO THE CARBOXYL TERMINUS
-arrestin and arrestin3, interact with high affinity and
stoichiometrically with clathrin, and we postulated that this
interaction mediates internalization of G protein-coupled receptors
(Goodman, O. B., Jr., Krupnick, J. G., Santini, F., Gurevich, V. V.,
Penn, R. B., Gagnon, A. W., Keen, J. H., and Benovic, J. L. (1996)
Nature 383, 447-450). In this study, we localized the
clathrin binding domain of arrestin3 using a variety of approaches.
Truncation mutagenesis demonstrated that the COOH-terminal half of
arrestin3 is required for clathrin interaction. Assessment of the
clathrin binding properties of various glutathione
S-transferase-arrestin3 fusion proteins indicated that the predominant
clathrin binding domain is contained within residues 367-385. Alanine
scanning mutagenesis further localized this domain to residues
371-379, and site-directed mutagenesis demonstrated the critical
importance of both hydrophobic (Leu-373, Ile-374, and Phe-376) and
acidic (Glu-375 and Glu-377) residues in the arrestin3/clathrin
interaction. These results are complementary to the observation that
hydrophobic and basic residues in clathrin are critical for its
interaction with nonvisual arrestins (Goodman, O. B., Jr., Krupnick,
J. G., Gurevich, V. V., Benovic, J. L., and Keen, J. H. (1997)
J. Biol. Chem. 272, 15017-15022). Lastly, an
arrestin3 mutant in which Leu-373, Ile-374, and Phe-376 were mutated to
Ala was significantly defective in its ability to promote 2-adrenergic receptor internalization in COS-1 cells
when compared with wild-type arrestin3. Taken together, these results
implicate a discrete region of arrestin3 in high affinity binding to
clathrin, an interaction critical for agonist-promoted internalization
of the 2-adrenergic receptor.
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J. Zhang, L. S. Barak, K. E. Winkler, M. G. Caron, and S. S. G. Ferguson A Central Role for beta -Arrestins and Clathrin-coated Vesicle-mediated Endocytosis in beta 2-Adrenergic Receptor Resensitization. DIFFERENTIAL REGULATION OF RECEPTOR RESENSITIZATION IN TWO DISTINCT CELL TYPES J. Biol. Chem., October 24, 1997; 272(43): 27005 - 27014. [Abstract] [Full Text] [PDF]
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O. B. Goodman Jr., J. G. Krupnick, V. V. Gurevich, J. L. Benovic, and J. H. Keen Arrestin/Clathrin Interaction. LOCALIZATION OF THE ARRESTIN BINDING LOCUS TO THE CLATHRIN TERMINAL DOMAIN J. Biol. Chem., June 6, 1997; 272(23): 15017 - 15022. [Abstract] [Full Text] [PDF]
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S. A. Laporte, R. H. Oakley, J. A. Holt, L. S. Barak, and M. G. Caron The Interaction of beta -Arrestin with the AP-2 Adaptor Is Required for the Clustering of beta 2-Adrenergic Receptor into Clathrin-coated Pits J. Biol. Chem., July 21, 2000; 275(30): 23120 - 23126. [Abstract] [Full Text] [PDF]
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A. Bhatnagar, D. L. Willins, J. A. Gray, J. Woods, J. L. Benovic, and B. L. Roth The Dynamin-dependent, Arrestin-independent Internalization of 5-Hydroxytryptamine 2A (5-HT2A) Serotonin Receptors Reveals Differential Sorting of Arrestins and 5-HT2A Receptors during Endocytosis J. Biol. Chem., March 9, 2001; 276(11): 8269 - 8277. [Abstract] [Full Text] [PDF]
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A. Fingerhut, K. von Figura, and S. Honing Binding of AP2 to Sorting Signals Is Modulated by AP2 Phosphorylation J. Biol. Chem., February 16, 2001; 276(8): 5476 - 5482. [Abstract] [Full Text] [PDF]
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W. Yang, C. G. Lo, T. Dispenza, and R. A. Cerione The Cdc42 Target ACK2 Directly Interacts with Clathrin and Influences Clathrin Assembly J. Biol. Chem., May 11, 2001; 276(20): 17468 - 17473. [Abstract] [Full Text] [PDF]
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T. Shiina, K. Arai, S. Tanabe, N. Yoshida, T. Haga, T. Nagao, and H. Kurose Clathrin Box in G Protein-coupled Receptor Kinase 2 J. Biol. Chem., August 24, 2001; 276(35): 33019 - 33026. [Abstract] [Full Text] [PDF]
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M. T. Drake and L. M. Traub Interaction of Two Structurally Distinct Sequence Types with the Clathrin Terminal Domain beta -Propeller J. Biol. Chem., July 27, 2001; 276(31): 28700 - 28709. [Abstract] [Full Text] [PDF]
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R. H. Oakley, S. A. Laporte, J. A. Holt, M. G. Caron, and L. S. Barak Differential Affinities of Visual Arrestin, beta Arrestin1, and beta Arrestin2 for G Protein-coupled Receptors Delineate Two Major Classes of Receptors J. Biol. Chem., May 26, 2000; 275(22): 17201 - 17210. [Abstract] [Full Text] [PDF]
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F. Santini, I. Gaidarov, and J. H. Keen G protein-coupled receptor/arrestin3 modulation of the endocytic machinery J. Cell Biol., February 18, 2002; 156(4): 665 - 676. [Abstract] [Full Text] [PDF]
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