Diverse cell surface protein ectodomains are shed by a system sensitive to metalloprotease inhibitors.
The extracellular domains of a diverse group of membrane proteins are shed in response to protein kinase C activators such as phorbol 12-myristate 13-acetate (PMA). The lack of sequence similarity in the cleavage sites suggests the involvement of many proteases of diverse specificity in this process. However, a mutant Chinese hamster ovary cell line recently isolated for being defective in PMA-activated shedding of the membrane-anchored growth factor transforming growth factor alpha precursor (proTGF-alpha) is concomitantly defective in the shedding of many other unrelated membrane proteins. Here we show that independent mutagenesis and selection experiments yield shedding mutants having the same recessive phenotype and belonging to the same genetic complementation group. Furthermore, two structurally distinct agents, TAPI-2 and 1,10-phenanthroline, which are known to inhibit metalloproteases, block PMA-activated shedding of proTGF-alpha, cell adhesion receptor L-selectin, interleukin 6 receptor alpha subunit, beta-amyloid precursor protein, and an entire set of anonymous Chinese hamster ovary cell surface proteins. Certain serine protease inhibitors prevent release of these proteins by interfering with their maturation and transport to the cell surface but do not inhibit ectodomain shedding from the cell surface. The results suggest the existence of a common system for membrane protein ectodomain shedding involving one or several proteolytic activities sensitive to metalloprotease inhibitors, whose ability to act can be disrupted by recessive mutations in a single gene.
Arribas
J
J
Cell Biology and Genetics Program, Memorial Sloan-Kettering Cancer Center, New York, New York 10021, USA.
Coodly
L
L
Vollmer
P
P
Kishimoto
T K
TK
Rose-John
S
S
Massagué
J
J
D016564:Amyloid beta-Protein Precursor / Q000378:metabolism; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D016466:CHO Cells; D002462:Cell Membrane / Q000378:metabolism; D006224:Cricetinae; D005816:Genetic Complementation Test; D006877:Hydroxamic Acids / Q000494:pharmacology*; D007700:Kinetics; D019041:L-Selectin / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism*; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors*; D016296:Mutagenesis; D010618:Phenanthrolines / Q000494:pharmacology*; D010641:Phenotype; D011480:Protease Inhibitors / Q000494:pharmacology*; D011498:Protein Precursors / Q000378:metabolism*; D018123:Receptors, Interleukin / Q000378:metabolism; D019947:Receptors, Interleukin-6; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D014162:Transfection; D016211:Transforming Growth Factor alpha / Q000096:biosynthesis / Q000378:metabolism*
D016564:Amyloid beta-Protein Precursor; D015703:Antigens, CD; D006877:Hydroxamic Acids; D008565:Membrane Proteins; D010618:Phenanthrolines; D011480:Protease Inhibitors; D011498:Protein Precursors; D018123:Receptors, Interleukin; D019947:Receptors, Interleukin-6; C099410:TAPI-2; D016211:Transforming Growth Factor alpha; C052268:protransforming growth factor alpha; D019041:L-Selectin; D008666:Metalloendopeptidases; D013755:Tetradecanoylphorbol Acetate; C025205:1,10-phenanthroline