Endocytosis of the growth hormone receptor (GHR) is regulated by the ubiquitin-conjugating system. A cytosolic 10 amino acid motif, referred to as the ubiquitin-dependent endocytosis (UbE) motif, is involved in the ubiquitination as well as in the endocytosis of the receptor. Proteins that are implicated in one of these processes have not been identified so far. Using a glutathione S-transferase (GST)-pulldown assay with a GST fusion protein encompassing the UbE motif of the GHR, a 35 kDa protein was purified. The protein was identified by MS as small glutamine-rich tetratricopeptide repeat (TPR)-containing protein (SGT). We found that GHR interacts with SGT. In vivo, both the precursor and the mature form of the receptor interacted with SGT. Inactivation of the ubiquitin-conjugating system did not affect the GHR–SGT interaction. Binding studies showed that the first TPR motif of SGT interacts with the UbE motif of the GHR. Taken together, these data show that SGT is a GHR-interacting protein, which binds independent of the ubiquitin-conjugating system.
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August 2003
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Research Article|
August 01 2003
Small glutamine-rich tetratricopeptide repeat-containing protein (SGT) interacts with the ubiquitin-dependent endocytosis (UbE) motif of the growth hormone receptor
Julia A. SCHANTL;
Julia A. SCHANTL
∗Department of Cell Biology and Institute of Biomembranes, University Medical Center, AZU G02.525, Heidelberglaan 100, 3584 CX Utrecht, The Netherlands
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Marcel ROZA;
Marcel ROZA
∗Department of Cell Biology and Institute of Biomembranes, University Medical Center, AZU G02.525, Heidelberglaan 100, 3584 CX Utrecht, The Netherlands
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Ad P. De JONG;
Ad P. De JONG
†Laboratory of Organic Analytical Chemistry, National Institute of Public Health and the Environment, Bilthoven, The Netherlands
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Ger J. STROUS
Ger J. STROUS
1
∗Department of Cell Biology and Institute of Biomembranes, University Medical Center, AZU G02.525, Heidelberglaan 100, 3584 CX Utrecht, The Netherlands
1To whom correspondence should be addressed (e-mail strous@med.uu.nl).
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Publisher: Portland Press Ltd
Received:
October 10 2002
Revision Received:
April 23 2003
Accepted:
May 08 2003
Accepted Manuscript online:
May 08 2003
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London ©2003
2003
Biochem J (2003) 373 (3): 855–863.
Article history
Received:
October 10 2002
Revision Received:
April 23 2003
Accepted:
May 08 2003
Accepted Manuscript online:
May 08 2003
Citation
Julia A. SCHANTL, Marcel ROZA, Ad P. De JONG, Ger J. STROUS; Small glutamine-rich tetratricopeptide repeat-containing protein (SGT) interacts with the ubiquitin-dependent endocytosis (UbE) motif of the growth hormone receptor. Biochem J 1 August 2003; 373 (3): 855–863. doi: https://doi.org/10.1042/bj20021591
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