Small glutamine-rich tetratricopeptide repeat-containing protein (SGT) interacts with the ubiquitin-dependent endocytosis (UbE) motif of the growth hormone receptor.
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.
Department of Cell Biology and Institute of Biomembranes, University Medical Center, AZU G02.525, Heidelberglaan 100, 3584 CX Utrecht, The Netherlands.
Julia A
JA
Schantl
Marcel
M
Roza
Ad P
AP
De Jong
Ger J
GJ
Strous
D020816:Amino Acid Motifs; D002352:Carrier Proteins; D002460:Cell Line; D004705:Endocytosis / Q000502:physiology*; D006801:Humans; D018832:Molecular Chaperones; D011506:Proteins / Q000378:metabolism*; D011986:Receptors, Somatotropin / Q000737:chemistry / Q000378:metabolism*; D025801:Ubiquitin / Q000378:metabolism*
D002352:Carrier Proteins; D018832:Molecular Chaperones; D011506:Proteins; D011986:Receptors, Somatotropin; C114575:SGTA protein, human; D025801:Ubiquitin