Human ADAM33: protein maturation and localization.
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.
Garlisi
Charles G
CG
Allergy, Schering-Plough Research Institute, 2015 Galloping Hill Road, Kenilworth, NJ 07033, USA. charles.garlisi@spcorp.com
Zou
Jun
J
Devito
Kristine E
KE
Tian
Fang
F
Zhu
Feng X
FX
Liu
Jianjun
J
Shah
Himanshu
H
Wan
Yuntao
Y
Motasim Billah
M
M
Egan
Robert W
RW
Umland
Shelby P
SP
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
D011480:Protease Inhibitors; D014020:Tissue Extracts; D000511:alpha-Macroglobulins; D051722:ADAM Proteins; C450441:ADAM33 protein, human; C499183:Adam33 protein, mouse; D008666:Metalloendopeptidases