Association of human transferrin receptor with GABARAP

Frank Green; Thomas O'Hare; Aaron Blackwell; Caroline A. Enns

doi:10.1016/S0014-5793(02)02655-8

Association of human transferrin receptor with GABARAP

Frank Green, Thomas O'Hare, Aaron Blackwell, Caroline A. Enns

Cell Developmental and Cancer Biology

Research output: Contribution to journal › Article › peer-review

52 Scopus citations

Abstract

A yeast two-hybrid screen identified a specific interaction between the cytoplasmic domain of transferrin receptor (TfR) and GABARAP, a 14 kDa protein that binds to the γ2 subunit of neuronal GABA_A receptors. The specificity of the TfR-GABARAP interaction was demonstrated by in vitro binding assays with purified proteins and by co-immunoprecipitation of GABARAP with endogenous TfR from HeLa cell lysates. Replacement of the YTRF internalization motif with ATRA within the cytoplasmic domain of TfR reduced interaction with GABARAP in the yeast two-hybrid screen and in vitro binding assays. The intracellular location of GABARAP using chimeric GABARAP-GFP showed that the majority of GABARAP was located in perinuclear vesicles. Our results show that GABARAP plays a more general role outside the confines of neuronal cells and GABA_A receptors.

Original language	English (US)
Pages (from-to)	101-106
Number of pages	6
Journal	FEBS Letters
Volume	518
Issue number	1-3
DOIs	https://doi.org/10.1016/S0014-5793(02)02655-8
State	Published - May 8 2002

Keywords

Endocytic motif
Protein trafficking
Transferrin receptor
γ-Aminobutyric acid type A receptor-associated protein

ASJC Scopus subject areas

Biophysics
Structural Biology
Biochemistry
Molecular Biology
Genetics
Cell Biology

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10.1016/S0014-5793(02)02655-8

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title = "Association of human transferrin receptor with GABARAP",

abstract = "A yeast two-hybrid screen identified a specific interaction between the cytoplasmic domain of transferrin receptor (TfR) and GABARAP, a 14 kDa protein that binds to the γ2 subunit of neuronal GABAA receptors. The specificity of the TfR-GABARAP interaction was demonstrated by in vitro binding assays with purified proteins and by co-immunoprecipitation of GABARAP with endogenous TfR from HeLa cell lysates. Replacement of the YTRF internalization motif with ATRA within the cytoplasmic domain of TfR reduced interaction with GABARAP in the yeast two-hybrid screen and in vitro binding assays. The intracellular location of GABARAP using chimeric GABARAP-GFP showed that the majority of GABARAP was located in perinuclear vesicles. Our results show that GABARAP plays a more general role outside the confines of neuronal cells and GABAA receptors.",

keywords = "Endocytic motif, Protein trafficking, Transferrin receptor, γ-Aminobutyric acid type A receptor-associated protein",

author = "Frank Green and Thomas O'Hare and Aaron Blackwell and Enns, {Caroline A.}",

note = "Funding Information: This work was supported by NIH DK 40608. We acknowledge Beatrice Pintea and James Little for technical assistance, Robin Warren and Anthony Williams for TfR constructs and thank Barb Root (Bristol Meyers-Squibb) for protein sequencing and Paul Howard, Dennis Glen and Stan Hollenberg for reagents and advice concerning the two-hybrid screen.",

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doi = "10.1016/S0014-5793(02)02655-8",

language = "English (US)",

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T1 - Association of human transferrin receptor with GABARAP

AU - Green, Frank

AU - O'Hare, Thomas

AU - Blackwell, Aaron

AU - Enns, Caroline A.

N1 - Funding Information: This work was supported by NIH DK 40608. We acknowledge Beatrice Pintea and James Little for technical assistance, Robin Warren and Anthony Williams for TfR constructs and thank Barb Root (Bristol Meyers-Squibb) for protein sequencing and Paul Howard, Dennis Glen and Stan Hollenberg for reagents and advice concerning the two-hybrid screen.

PY - 2002/5/8

Y1 - 2002/5/8

N2 - A yeast two-hybrid screen identified a specific interaction between the cytoplasmic domain of transferrin receptor (TfR) and GABARAP, a 14 kDa protein that binds to the γ2 subunit of neuronal GABAA receptors. The specificity of the TfR-GABARAP interaction was demonstrated by in vitro binding assays with purified proteins and by co-immunoprecipitation of GABARAP with endogenous TfR from HeLa cell lysates. Replacement of the YTRF internalization motif with ATRA within the cytoplasmic domain of TfR reduced interaction with GABARAP in the yeast two-hybrid screen and in vitro binding assays. The intracellular location of GABARAP using chimeric GABARAP-GFP showed that the majority of GABARAP was located in perinuclear vesicles. Our results show that GABARAP plays a more general role outside the confines of neuronal cells and GABAA receptors.

AB - A yeast two-hybrid screen identified a specific interaction between the cytoplasmic domain of transferrin receptor (TfR) and GABARAP, a 14 kDa protein that binds to the γ2 subunit of neuronal GABAA receptors. The specificity of the TfR-GABARAP interaction was demonstrated by in vitro binding assays with purified proteins and by co-immunoprecipitation of GABARAP with endogenous TfR from HeLa cell lysates. Replacement of the YTRF internalization motif with ATRA within the cytoplasmic domain of TfR reduced interaction with GABARAP in the yeast two-hybrid screen and in vitro binding assays. The intracellular location of GABARAP using chimeric GABARAP-GFP showed that the majority of GABARAP was located in perinuclear vesicles. Our results show that GABARAP plays a more general role outside the confines of neuronal cells and GABAA receptors.

KW - Endocytic motif

KW - Protein trafficking

KW - Transferrin receptor

KW - γ-Aminobutyric acid type A receptor-associated protein

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Association of human transferrin receptor with GABARAP

Abstract

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ASJC Scopus subject areas

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