title: Human ADAM33: protein maturation and localization.
abstract: ADAM33 (a disintegrin and metalloprotease) was recently found to be a novel asthma susceptibility gene. Domain-specific antibodies were used to study its expression and processing. When the pro-domain and catalytic domain were expressed by a stable-transfected cell line, the pro-domain was removed by cleavage within a putative furin cleavage site. The catalytic domain was active in an alpha(2)-macroglobulin complex formation assay and mutation of the catalytic site glutamic acid (E346A) eliminated activity. In transient transfections using the full-length protein, a pro-form and mature form were detectable and alternate glycosylation was demonstrated at sites within the catalytic domain. ADAM33 was detected on the cell surface, with the majority of protein detected intracellularly. The E346A mutation had no significant effect on protein processing. Endogenous ADAM33 was detected in bronchus tissue, bronchial smooth muscle cells, and MRC-5 fibroblasts, consistent with a role in the pathophysiology of asthma.
authors.affiliation: Allergy, Schering-Plough Research Institute, 2015 Galloping Hill Road, Kenilworth, NJ 07033, USA. charles.garlisi@spcorp.com
authors.forename: Charles G
authors.initials: CG
authors.lastname: Garlisi
authors.forename: Jun
authors.initials: J
authors.lastname: Zou
authors.forename: Kristine E
authors.initials: KE
authors.lastname: Devito
authors.forename: Fang
authors.initials: F
authors.lastname: Tian
authors.forename: Feng X
authors.initials: FX
authors.lastname: Zhu
authors.forename: Jianjun
authors.initials: J
authors.lastname: Liu
authors.forename: Himanshu
authors.initials: H
authors.lastname: Shah
authors.forename: Yuntao
authors.initials: Y
authors.lastname: Wan
authors.forename: M
authors.initials: M
authors.lastname: Motasim Billah
authors.forename: Robert W
authors.initials: RW
authors.lastname: Egan
authors.forename: Shelby P
authors.initials: SP
authors.lastname: Umland
mesh_terms: D051722:ADAM Proteins; D000818:Animals; D020134:Catalytic Domain; D002460:Cell Line; D006031:Glycosylation; D006801:Humans; D008666:Metalloendopeptidases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D009154:Mutation; D011110:Polymorphism, Genetic; D011480:Protease Inhibitors / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D014018:Tissue Distribution; D014020:Tissue Extracts / Q000378:metabolism; D014162:Transfection; D000511:alpha-Macroglobulins / Q000378:metabolism
chemical_list: D011480:Protease Inhibitors; D014020:Tissue Extracts; D000511:alpha-Macroglobulins; D051722:ADAM Proteins; C450441:ADAM33 protein, human; C499183:Adam33 protein, mouse; D008666:Metalloendopeptidases