Biochemical and Pharmacological Criteria Define Two Shedding Activities for TRANCE/OPGL That Are Distinct from the Tumor Necrosis Factor α Convertase

A number of structurally and functionally diverse membrane proteins are released from the plasma membrane in a process termed protein ectodomain shedding. Ectodomain shedding may activate or inactivate a substrate or change its properties, such as converting a juxtacrine into a paracrine signaling molecule. Here we have characterized the activities involved in protein ectodomain shedding of the tumor necrosis factor family member TRANCE/OPGL in different cell types. The criteria used to evaluate these activities include (a) cleavage site usage, (b) response to activators and inhibitors of intracellular signaling pathways, and (c) sensitivity to tissue inhibitors of metalloproteases (TIMPs). At least two TRANCE shedding activities emerged, both of which are distinct from the tumor necrosis factor α convertase. One of the TRANCE sheddases is induced by the tyrosine phosphatase inhibitor pervanadate but not by phorbol esters, whereas the other is refractory to both of these stimuli. Furthermore, the pervanadate-regulated sheddase activity is sensitive to TIMP-2 but not TIMP-1, which is consistent with the properties of a membrane type matrix metalloprotease. This study provides insights into the properties of different activities involved in protein ectodomain shedding and has implications for the functional regulation of TRANCE by potentially more than one protease.