Shedding of human thyrotropin receptor ectodomain. Involvement of a matrix metalloprotease.
The thyrotropin (TSH) receptor in human thyroid glands has been shown to be cleaved into an extracellular alpha subunit and a transmembrane beta subunit held together by disulfide bridges. An excess of the latter component relative to the former suggested the shedding of the ectodomain. Indeed we observed such a shedding in cultures of human thyrocytes and permanently transfected L or Chinese hamster ovary cells. The shedding was increased by inhibitors of endocytosis, recycling, and lysosomal degradation, suggesting that it was dependent on receptor residency at the cell surface. It was slightly increased by TSH and phorbol esters, whereas forskolin and 8-bromo-cyclic AMP were without effect. Decreasing the serum concentration in cell culture medium enhanced the shedding by an unknown mechanism. The shedding of the TSH receptor alpha domain is the consequence of two events: cleavage of the receptor into alpha and beta subunits and reduction of the disulfide bridge(s). The complete inhibition of soluble TSH receptor shedding by the specific inhibitor BB-2116 indicated that the cleavage reaction is catalyzed probably at the cell surface by a matrix metalloprotease. This shedding mechanism may be responsible for the presence of soluble TSH receptor alpha subunit in human circulation.
Couet
J
J
Unité de Recherches Hormones et Reproduction, Institut National de la Santé et de la Recherche Médicale, Unité 135, Le Kremlin-Bicêtre, France.
Sar
S
S
Jolivet
A
A
Hai
M T
MT
Milgrom
E
E
Misrahi
M
M
D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology; D000818:Animals; D016466:CHO Cells; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D005576:Colforsin / Q000494:pharmacology; D006224:Cricetinae; D004705:Endocytosis; D005109:Extracellular Matrix / Q000201:enzymology*; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D007700:Kinetics; D007739:L Cells; D008247:Lysosomes / Q000378:metabolism; D046911:Macromolecular Substances; D008666:Metalloendopeptidases / Q000378:metabolism*; D051379:Mice; D011480:Protease Inhibitors / Q000494:pharmacology; D011989:Receptors, Thyrotropin / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013961:Thyroid Gland / Q000378:metabolism*; D014162:Transfection
C088670:BB 2116; D006877:Hydroxamic Acids; D046911:Macromolecular Substances; D011480:Protease Inhibitors; D011989:Receptors, Thyrotropin; D011994:Recombinant Proteins; D005576:Colforsin; D015124:8-Bromo Cyclic Adenosine Monophosphate; D008666:Metalloendopeptidases; D013755:Tetradecanoylphorbol Acetate