ADAM28 a member of the ADAM (a disintegrin and metalloproteinase) gene family is over-expressed by carcinoma cells and the expression correlates with carcinoma cell proliferation and progression in human lung and breast carcinomas However information about substrates of ADAM28 is limited We screened interacting molecules of ADAM28 in human lung cDNA library by yeast two-hybrid system and identified connective tissue growth factor (CTGF) Binding of CTGF to proADAM28 was demonstrated by yeast two-hybrid assay and protein binding assay ADAM28 cleaved CTGF in dose- and time-dependent manners at the Ala(181)-Tyr(182) and Asp(191)-Pro(192)bonds in the hinge region of the molecule ADAM28 selectively digested CTGF in the complex of CTGF and vascular endothelial growth factor(165) (VEGF(165)) releasing biologically active VEGF(165) from the complex RT-PCR and immunohistochemical analyses demonstrated that ADAM28 CTGF and VEGF are commonly co-expressed in the breast carcinoma tissues These data provide the first evidence that CTGF is a novel substrate of ADAM28 and suggest that ADAM28 may promote VEGF(165)-induced angiogenesis in the breast carcinomas by the CTGF digestion in the CTGF/VEGF(165) complex (C) 2010 Elsevier Inc All rights reserved