Abstract
ABCA3 mutations cause fatal surfactant deficiency and interstitial lung disease. ABCA3 protein is a lipid transporter indispensible for surfactant biogenesis and storage in lamellar bodies (LB). The protein folds in endoplasmic reticulum and is glycosylated in Golgi en route to the membrane of mature LB and their precursor multivesicular bodies (MVB). In immunoblots, C-terminally labeled ABCA3 appears as two protein bands of 150 and 190 kDa. Using N- and C-terminal protein tags and hindering ABCA3 processing we show that the 150 kDa protein represents the mature ABCA3 whose N-terminus is cleaved by a cysteine protease inside MVB/LB.
Copyright © 2010 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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ATP-Binding Cassette Transporters / chemistry
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ATP-Binding Cassette Transporters / genetics
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ATP-Binding Cassette Transporters / metabolism*
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Cell Line, Tumor
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Cysteine Proteinase Inhibitors / pharmacology
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Endothelium, Vascular / metabolism
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Glycosylation
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Golgi Apparatus / metabolism
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Humans
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Immunoblotting
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Leucine / analogs & derivatives
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Leucine / pharmacology
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Luminescent Proteins / genetics
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Luminescent Proteins / metabolism
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Lysosomal Membrane Proteins / metabolism*
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Microscopy, Fluorescence
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Molecular Weight
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Multivesicular Bodies / metabolism*
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Neoplasm Proteins / metabolism*
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Pepstatins / pharmacology
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Protein Folding
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Protein Processing, Post-Translational / drug effects
Substances
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ABCA3 protein, human
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ATP-Binding Cassette Transporters
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Cysteine Proteinase Inhibitors
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LAMP3 protein, human
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Luminescent Proteins
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Lysosomal Membrane Proteins
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Neoplasm Proteins
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Pepstatins
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Leucine
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E 64
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pepstatin