Identification and characterization of a novel ABCA3 mutation. Physiol Genomics 2010 Jan 08;40(2):94-9
Date
10/29/2009Pubmed ID
19861431DOI
10.1152/physiolgenomics.00123.2009Scopus ID
2-s2.0-73949109195 (requires institutional sign-in at Scopus site) 17 CitationsAbstract
Mutations in the gene coding for ATP-binding cassette protein A3 (ABCA3) are recognized as a genetic cause of lung disease of varying severity. Characterization of a number of mutant ABCA3 proteins has demonstrated that the mutations generally affect intracellular localization or the ability of the protein to hydrolyze ATP. A novel heterozygous mutation that results in the substitution of cysteine for arginine at amino acid 295 in ABCA3 was identified in a premature infant with chronic respiratory insufficiency and abnormal lamellar bodies. Sequencing of DNA performed in study participants demonstrated that this was a mutation and not a common variant. Plasmid vectors containing ABCA3 with the identified novel mutation tagged with green fluorescent protein on the carboxy terminus were generated. The effect of the mutation on protein function was characterized by examining the glycosylation state of the mutant protein in transiently transfected HEK293 cells and by examining ATP hydrolysis activity of the mutant protein with a vanadate-induced nucleotide trapping assay in stably transfected HEK293 cells. The ABCA3 protein containing the R295C mutation undergoes normal glycosylation and intracellular localization but has dramatically reduced ATP hydrolysis activity (12% of wild type). The identification of one copy of this novel mutation in a premature infant with chronic respiratory insufficiency suggests that ABCA3 haploinsufficiency together with lung prematurity may result in more severe, or more prolonged, respiratory failure.
Author List
Park SK, Amos L, Rao A, Quasney MW, Matsumura Y, Inagaki N, Dahmer MKAuthor
Louella B. Amos MD Associate Professor in the Pediatrics department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
ATP-Binding Cassette TransportersAdenosine Triphosphate
Amino Acid Sequence
Cells, Cultured
Glycosylation
Humans
Hydrolysis
Molecular Sequence Data
Mutation
Transfection
Young Adult
