DOI: 10.1046/j.1432-1327.1999.00549.x. Dynamin and Rab5 regulate GRK2‐dependent internalization of dopamine D2 receptors

Dynamin and Rab5 regulate GRK2‐dependent internalization of dopamine D2 receptors

10.1046/j.1432-1327.1999.00549.x

Crossref journal-article

Wiley

European Journal of Biochemistry (311)

Abstract

Dopamine D2 receptors (D2Rs; short form, which is one of the alternative splicing variants) expressed in COS‐7 cells are internalized in an agonist‐dependent manner only when G protein‐coupled receptor kinase 2 (GRK2) is coexpressed [Ito, K., Haga, T., Lameh, J. & Sadée, W., (1999) Eur. J. Biochem.260, 112–119]. We have examined the effects of coexpression of dynamin, a small molecular mass GTP‐binding protein, rab5A, and their mutants on the internalization of D2Rs in the presence of both dopamine (10 or 100 µm) and GRK2. The rate and extent of D2R internalization was increased or decreased by coexpression of dynamin I or a dominant‐negative form of dynamin I (dynamin I K44E), respectively. The effects of coexpressing these two dynamins were more prominent at 10 µm dopamine than at 100 µm. In the presence of 10 µm dopamine, internalization of D2R was completely suppressed when dynamin I K44E was coexpressed, and the half‐life (t½) of D2R internalization decreased relative to cells not expressing dynamin from 82 to 29 min when dynamin I was coexpressed. Internalization of D2Rs was facilitated or suppressed by coexpression of a constitutively active form of rab5A (rab5A Q79L) or a dominant‐negative form of rab5A (rab5A S34N), respectively. The t½ of D2R internalization at 10 µm dopamine decreased from 82 to 16 min in cells coexpressing rab5A Q79L. The effect of coexpression of rab5A S34N was more apparent at 100 µm dopamine than at 10 µm; the t½ of D2R internalization at 100 µm dopamine increased from 20 to 56 min and the proportion of internalized D2Rs after 120 min decreased from 53 to 28%. These results indicate that the internalization of D2Rs is dependent on the action of dynamin as well as GRK2, and is regulated by the action of rab5A.

Bibliography

Iwata, K., Ito, K., Fukuzaki, A., Inaki, K., & Haga, T. (1999). Dynamin and Rab5 regulate GRK2âdependent internalization of dopamine D2 receptors. European Journal of Biochemistry, 263(2), 596â602. Portico.

Authors 5

Ken Iwata (first)
Kazumitsu Ito (additional)
Atsushi Fukuzaki (additional)
Kouichiro Inaki (additional)
Tatsuya Haga (additional)

References 37 Referenced 85

10.1016/S0165-6147(97)01091-2
10.1074/jbc.273.30.18677
10.1146/annurev.pharmtox.38.1.289
10.1016/S0021-9258(18)31665-X / J. Biol. Chem. / Sequestration of muscarinic acetylcholine receptor m2 subtypes. Facilitation by G protein‐coupled receptor kinase (GRK2) and attenuation by a dominant‐negative mutant of GRK2. by Tsuga H. (1994)
10.1093/emboj/16.15.4606
10.1074/jbc.270.42.24782
10.1126/science.271.5247.363
10.1038/383447a0
10.1038/374190a0
10.1074/jbc.271.31.18302
10.1074/jbc.272.43.27124
10.1074/jbc.273.21.12967
10.1074/jbc.273.20.12155
10.1074/jbc.270.48.29004
{'key': 'e_1_2_7_16_2', 'first-page': '1218', 'article-title': 'Sequestration of human muscarinic acetylcholine receptor hm1‐hm5 subtypes: effect of G protein‐coupled receptor kinases GRK2, GRK4, GRK5 and GRK6.', 'volume': '284', 'author': 'Tsuga H.', 'year': '1998', 'journal-title': 'J. Pharmacol. Exp. Ther.'} / J. Pharmacol. Exp. Ther. / Sequestration of human muscarinic acetylcholine receptor hm1‐hm5 subtypes: effect of G protein‐coupled receptor kinases GRK2, GRK4, GRK5 and GRK6. by Tsuga H. (1998)
10.1016/0092-8674(87)90623-4
10.1038/333370a0
{'key': 'e_1_2_7_19_2', 'first-page': '192', 'article-title': 'Mutations of the human β2‐adrenergic receptor that impair coupling to Gs interfere with receptor down‐regulation but not sequestration.', 'volume': '39', 'author': 'Campbell P.T.', 'year': '1991', 'journal-title': 'Mol. Pharmacol.'} / Mol. Pharmacol. / Mutations of the human β2‐adrenergic receptor that impair coupling to Gs interfere with receptor down‐regulation but not sequestration. by Campbell P.T. (1991)
10.1074/jbc.273.16.9878
10.1074/jbc.273.13.7637
10.1074/jbc.272.38.23682
10.1074/jbc.273.39.24987
10.1046/j.1432-1327.1999.00125.x
10.1016/0092-8674(90)90369-P
10.1016/S0021-9258(20)80550-X / J. Biol. Chem. / Structure‐function relationship of the small GTPase rab5. by Li G. (1993)
10.1074/jbc.270.19.11257
10.1002/j.1460-2075.1994.tb06381.x
10.1242/jcs.108.9.2983 / J. Cell Sci. / Ligand‐stimulated β2‐adrenergic receptor internalization via the constitutive endocytic pathway into rab5‐containing endosomes. by Moore R.H. (1995)
10.1093/nar/18.17.5322
10.1074/jbc.271.51.33067
{'key': 'e_1_2_7_32_2', 'first-page': '560', 'article-title': 'Sequestration of the short and long isoforms of dopamine D2 receptors expressed in Chinese hamster ovary cells.', 'volume': '49', 'author': 'Itokawa M.', 'year': '1996', 'journal-title': 'Mol. Pharmacol.'} / Mol. Pharmacol. / Sequestration of the short and long isoforms of dopamine D2 receptors expressed in Chinese hamster ovary cells. by Itokawa M. (1996)
{'key': 'e_1_2_7_33_2', 'first-page': '378', 'article-title': 'Ligand binding properties of muscarinic acetylcholine receptor subtypes (m1‐m5) expressed in baculovirus‐infected insect cells.', 'volume': '274', 'author': 'Guo Z.D.', 'year': '1995', 'journal-title': 'J. Pharmacol. Exp. Ther.'} / J. Pharmacol. Exp. Ther. / Ligand binding properties of muscarinic acetylcholine receptor subtypes (m1‐m5) expressed in baculovirus‐infected insect cells. by Guo Z.D. (1995)
10.1124/mol.51.5.800 / Mol. Pharmacol. / Synergistic regulation of β2‐adrenergic receptor sequestration: intracellular complement of β‐adrenergic receptor kinase and β‐arrestin determine kinetics of internalization. by Menard L. (1997)
10.1073/pnas.95.12.7157
10.1016/S0021-9258(17)36671-1 / J. Biol. Chem. / Structural features of the GTP‐binding defective Rab5 mutants required for their inhibitory activity on endocytosis. by Li G. (1994)
10.1016/S0092-8674(00)80380-3
10.1074/jbc.273.38.24592

Dates

Type	When
Created	22 years, 5 months ago (March 11, 2003, 12:27 p.m.)
Deposited	1 year, 10 months ago (Oct. 16, 2023, 10:59 a.m.)
Indexed	1 week, 6 days ago (Aug. 23, 2025, 12:58 a.m.)
Issued	26 years, 1 month ago (July 15, 1999)
Published	26 years, 1 month ago (July 15, 1999)
Published Online	23 years, 8 months ago (Dec. 25, 2001)
Published Print	26 years, 1 month ago (July 15, 1999)

Funders 0

None

BibTeX

@article{Iwata_1999, title={Dynamin and Rab5 regulate GRK2‐dependent internalization of dopamine D2 receptors}, volume={263}, ISSN={1432-1033}, url={http://dx.doi.org/10.1046/j.1432-1327.1999.00549.x}, DOI={10.1046/j.1432-1327.1999.00549.x}, number={2}, journal={European Journal of Biochemistry}, publisher={Wiley}, author={Iwata, Ken and Ito, Kazumitsu and Fukuzaki, Atsushi and Inaki, Kouichiro and Haga, Tatsuya}, year={1999}, month=jul, pages={596–602} }

JSON

{
  "indexed": {
    "date-parts": [
      [
        2025,
        8,
        23
      ]
    ],
    "date-time": "2025-08-23T04:58:53Z",
    "timestamp": 1755925133311
  },
  "reference-count": 37,
  "publisher": "Wiley",
  "issue": "2",
  "license": [
    {
      "start": {
        "date-parts": [
          [
            2001,
            12,
            25
          ]
        ],
        "date-time": "2001-12-25T00:00:00Z",
        "timestamp": 1009238400000
      },
      "content-version": "vor",
      "delay-in-days": 894,
      "URL": "http://onlinelibrary.wiley.com/termsAndConditions#vor"
    }
  ],
  "content-domain": {
    "domain": [],
    "crossmark-restriction": false
  },
  "published-print": {
    "date-parts": [
      [
        1999,
        7,
        15
      ]
    ]
  },
  "abstract": "<jats:p>Dopamine D2 receptors (D2Rs; short form, which is one of the alternative splicing variants) expressed in COS\u20107 cells are internalized in an agonist\u2010dependent manner only when G\u2003protein\u2010coupled receptor kinase\u20032 (GRK2) is coexpressed [Ito, K., Haga, T., Lameh, J. &amp; Sad\u00e9e, W., (1999) <jats:italic>Eur. J. Biochem.</jats:italic><jats:bold>260</jats:bold>, 112\u2013119]. We have examined the effects of coexpression of dynamin, a small molecular mass GTP\u2010binding protein, rab5A, and their mutants on the internalization of D2Rs in the presence of both dopamine (10 or 100\u2003\u00b5<jats:sc>m</jats:sc>) and GRK2. The rate and extent of D2R internalization was increased or decreased by coexpression of dynamin\u2003I or a dominant\u2010negative form of dynamin\u2003I (dynamin\u2003I K44E), respectively. The effects of coexpressing these two dynamins were more prominent at 10\u2003\u00b5<jats:sc>m</jats:sc> dopamine than at 100\u2003\u00b5<jats:sc>m</jats:sc>. In the presence of 10\u2003\u00b5<jats:sc>m</jats:sc> dopamine, internalization of D2R was completely suppressed when dynamin\u2003I K44E was coexpressed, and the half\u2010life (<jats:italic>t</jats:italic><jats:sub>\u00bd</jats:sub>) of D2R internalization decreased relative to cells not expressing dynamin from 82 to 29\u2003min when dynamin\u2003I was coexpressed. Internalization of D2Rs was facilitated or suppressed by coexpression of a constitutively active form of rab5A (rab5A Q79L) or a dominant\u2010negative form of rab5A (rab5A S34N), respectively. The <jats:italic>t</jats:italic><jats:sub>\u00bd</jats:sub> of D2R internalization at 10\u2003\u00b5<jats:sc>m</jats:sc> dopamine decreased from 82 to 16\u2003min in cells coexpressing rab5A Q79L. The effect of coexpression of rab5A S34N was more apparent at 100\u2003\u00b5<jats:sc>m</jats:sc> dopamine than at 10\u2003\u00b5<jats:sc>m</jats:sc>; the <jats:italic>t</jats:italic><jats:sub>\u00bd</jats:sub> of D2R internalization at 100\u2003\u00b5<jats:sc>m</jats:sc> dopamine increased from 20 to 56\u2003min and the proportion of internalized D2Rs after 120\u2003min decreased from 53 to 28%. These results indicate that the internalization of D2Rs is dependent on the action of dynamin as well as GRK2, and is regulated by the action of rab5A.</jats:p>",
  "DOI": "10.1046/j.1432-1327.1999.00549.x",
  "type": "journal-article",
  "created": {
    "date-parts": [
      [
        2003,
        3,
        11
      ]
    ],
    "date-time": "2003-03-11T17:27:02Z",
    "timestamp": 1047403622000
  },
  "page": "596-602",
  "source": "Crossref",
  "is-referenced-by-count": 85,
  "title": "Dynamin and Rab5 regulate GRK2\u2010dependent internalization of dopamine D2 receptors",
  "prefix": "10.1111",
  "volume": "263",
  "author": [
    {
      "given": "Ken",
      "family": "Iwata",
      "sequence": "first",
      "affiliation": []
    },
    {
      "given": "Kazumitsu",
      "family": "Ito",
      "sequence": "additional",
      "affiliation": []
    },
    {
      "given": "Atsushi",
      "family": "Fukuzaki",
      "sequence": "additional",
      "affiliation": []
    },
    {
      "given": "Kouichiro",
      "family": "Inaki",
      "sequence": "additional",
      "affiliation": []
    },
    {
      "given": "Tatsuya",
      "family": "Haga",
      "sequence": "additional",
      "affiliation": []
    }
  ],
  "member": "311",
  "published-online": {
    "date-parts": [
      [
        2001,
        12,
        25
      ]
    ]
  },
  "reference": [
    {
      "key": "e_1_2_7_2_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0165-6147(97)01091-2"
    },
    {
      "key": "e_1_2_7_3_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.273.30.18677"
    },
    {
      "key": "e_1_2_7_4_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1146/annurev.pharmtox.38.1.289"
    },
    {
      "key": "e_1_2_7_5_2",
      "doi-asserted-by": "crossref",
      "first-page": "32522",
      "DOI": "10.1016/S0021-9258(18)31665-X",
      "article-title": "Sequestration of muscarinic acetylcholine receptor m2 subtypes. Facilitation by G protein\u2010coupled receptor kinase (GRK2) and attenuation by a dominant\u2010negative mutant of GRK2.",
      "volume": "269",
      "author": "Tsuga H.",
      "year": "1994",
      "journal-title": "J. Biol. Chem."
    },
    {
      "key": "e_1_2_7_6_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/emboj/16.15.4606"
    },
    {
      "key": "e_1_2_7_7_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.270.42.24782"
    },
    {
      "key": "e_1_2_7_8_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.271.5247.363"
    },
    {
      "key": "e_1_2_7_9_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/383447a0"
    },
    {
      "key": "e_1_2_7_10_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/374190a0"
    },
    {
      "key": "e_1_2_7_11_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.271.31.18302"
    },
    {
      "key": "e_1_2_7_12_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.272.43.27124"
    },
    {
      "key": "e_1_2_7_13_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.273.21.12967"
    },
    {
      "key": "e_1_2_7_14_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.273.20.12155"
    },
    {
      "key": "e_1_2_7_15_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.270.48.29004"
    },
    {
      "key": "e_1_2_7_16_2",
      "first-page": "1218",
      "article-title": "Sequestration of human muscarinic acetylcholine receptor hm1\u2010hm5 subtypes: effect of G protein\u2010coupled receptor kinases GRK2, GRK4, GRK5 and GRK6.",
      "volume": "284",
      "author": "Tsuga H.",
      "year": "1998",
      "journal-title": "J. Pharmacol. Exp. Ther."
    },
    {
      "key": "e_1_2_7_17_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0092-8674(87)90623-4"
    },
    {
      "key": "e_1_2_7_18_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/333370a0"
    },
    {
      "key": "e_1_2_7_19_2",
      "first-page": "192",
      "article-title": "Mutations of the human \u03b22\u2010adrenergic receptor that impair coupling to Gs interfere with receptor down\u2010regulation but not sequestration.",
      "volume": "39",
      "author": "Campbell P.T.",
      "year": "1991",
      "journal-title": "Mol. Pharmacol."
    },
    {
      "key": "e_1_2_7_20_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.273.16.9878"
    },
    {
      "key": "e_1_2_7_21_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.273.13.7637"
    },
    {
      "key": "e_1_2_7_22_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.272.38.23682"
    },
    {
      "key": "e_1_2_7_23_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.273.39.24987"
    },
    {
      "key": "e_1_2_7_24_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1046/j.1432-1327.1999.00125.x"
    },
    {
      "key": "e_1_2_7_25_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0092-8674(90)90369-P"
    },
    {
      "key": "e_1_2_7_26_2",
      "doi-asserted-by": "crossref",
      "first-page": "24475",
      "DOI": "10.1016/S0021-9258(20)80550-X",
      "article-title": "Structure\u2010function relationship of the small GTPase rab5.",
      "volume": "268",
      "author": "Li G.",
      "year": "1993",
      "journal-title": "J. Biol. Chem."
    },
    {
      "key": "e_1_2_7_27_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.270.19.11257"
    },
    {
      "key": "e_1_2_7_28_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1002/j.1460-2075.1994.tb06381.x"
    },
    {
      "key": "e_1_2_7_29_2",
      "doi-asserted-by": "crossref",
      "first-page": "2983",
      "DOI": "10.1242/jcs.108.9.2983",
      "article-title": "Ligand\u2010stimulated \u03b22\u2010adrenergic receptor internalization via the constitutive endocytic pathway into rab5\u2010containing endosomes.",
      "volume": "108",
      "author": "Moore R.H.",
      "year": "1995",
      "journal-title": "J. Cell Sci."
    },
    {
      "key": "e_1_2_7_30_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/nar/18.17.5322"
    },
    {
      "key": "e_1_2_7_31_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.271.51.33067"
    },
    {
      "key": "e_1_2_7_32_2",
      "first-page": "560",
      "article-title": "Sequestration of the short and long isoforms of dopamine D2 receptors expressed in Chinese hamster ovary cells.",
      "volume": "49",
      "author": "Itokawa M.",
      "year": "1996",
      "journal-title": "Mol. Pharmacol."
    },
    {
      "key": "e_1_2_7_33_2",
      "first-page": "378",
      "article-title": "Ligand binding properties of muscarinic acetylcholine receptor subtypes (m1\u2010m5) expressed in baculovirus\u2010infected insect cells.",
      "volume": "274",
      "author": "Guo Z.D.",
      "year": "1995",
      "journal-title": "J. Pharmacol. Exp. Ther."
    },
    {
      "key": "e_1_2_7_34_2",
      "doi-asserted-by": "crossref",
      "first-page": "800",
      "DOI": "10.1124/mol.51.5.800",
      "article-title": "Synergistic regulation of \u03b22\u2010adrenergic receptor sequestration: intracellular complement of \u03b2\u2010adrenergic receptor kinase and \u03b2\u2010arrestin determine kinetics of internalization.",
      "volume": "51",
      "author": "Menard L.",
      "year": "1997",
      "journal-title": "Mol. Pharmacol."
    },
    {
      "key": "e_1_2_7_35_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.95.12.7157"
    },
    {
      "key": "e_1_2_7_36_2",
      "doi-asserted-by": "crossref",
      "first-page": "14631",
      "DOI": "10.1016/S0021-9258(17)36671-1",
      "article-title": "Structural features of the GTP\u2010binding defective Rab5 mutants required for their inhibitory activity on endocytosis.",
      "volume": "269",
      "author": "Li G.",
      "year": "1994",
      "journal-title": "J. Biol. Chem."
    },
    {
      "key": "e_1_2_7_37_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0092-8674(00)80380-3"
    },
    {
      "key": "e_1_2_7_38_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.273.38.24592"
    }
  ],
  "container-title": "European Journal of Biochemistry",
  "original-title": [],
  "language": "en",
  "link": [
    {
      "URL": "https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1046%2Fj.1432-1327.1999.00549.x",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "text-mining"
    },
    {
      "URL": "https://febs.onlinelibrary.wiley.com/doi/pdf/10.1046/j.1432-1327.1999.00549.x",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "similarity-checking"
    }
  ],
  "deposited": {
    "date-parts": [
      [
        2023,
        10,
        16
      ]
    ],
    "date-time": "2023-10-16T14:59:23Z",
    "timestamp": 1697468363000
  },
  "score": 1,
  "resource": {
    "primary": {
      "URL": "https://febs.onlinelibrary.wiley.com/doi/10.1046/j.1432-1327.1999.00549.x"
    }
  },
  "subtitle": [],
  "short-title": [],
  "issued": {
    "date-parts": [
      [
        1999,
        7,
        15
      ]
    ]
  },
  "references-count": 37,
  "journal-issue": {
    "issue": "2",
    "published-print": {
      "date-parts": [
        [
          1999,
          7,
          15
        ]
      ]
    }
  },
  "alternative-id": [
    "10.1046/j.1432-1327.1999.00549.x"
  ],
  "URL": "http://dx.doi.org/10.1046/j.1432-1327.1999.00549.x",
  "relation": {},
  "ISSN": [
    "0014-2956",
    "1432-1033"
  ],
  "subject": [],
  "container-title-short": "European Journal of Biochemistry",
  "published": {
    "date-parts": [
      [
        1999,
        7,
        15
      ]
    ]
  }
}