ABCMdb

PMID: 23543682

Lyssenko NN, Nickel M, Tang C, Phillips MC
Factors controlling nascent high-density lipoprotein particle heterogeneity: ATP-binding cassette transporter A1 activity and cell lipid and apolipoprotein AI availability.
FASEB J. 2013 Jul;27(7):2880-92. doi: 10.1096/fj.12-216564. Epub 2013 Mar 29., [PubMed]

Sentences

No. Mutations Sentence Comment

37 ABCA1 p.Trp590Ser ABCA1 p.Cys1477Arg MATERIALS AND METHODS Cell culture and biogenesis of nascent HDL BHK-ABCA1, BHK-ABCA1(W590S), and BHK-ABCA1(C1477R) cells expressing human wild-type or mutant ABCA1 have been previously described (36, 37). Login to comment 166 ABCA1 p.Trp590Ser ABCA1 p.Cys1477Arg Mutations in ABCA1 that compromise its functionality shift nascent HDL production to the smaller particle species W590S and C1477R are naturally occurring mutations in human ABCA1 that impair its functionality and cause Tangier disease (49) but do not affect its subcellular localization or expression levels (37). Login to comment 167 ABCA1 p.Trp590Ser ABCA1 p.Cys1477Arg To determine how deficiency in ABCA1 functionality affects size heterogeneity of nascent HDL particles, BHK-ABCA1, BHK-ABCA1(W590S), and BHK-ABCA1(C1477R) cells were plated at the same density, labeled with [3 H]cholesterol, induced to express ABCA1 at the maximum level, and exposed to a high concentration of human lipid-free apoAI, followed by the regular procedure to collect and analyze nascent HDL via gel filtration. Login to comment 168 ABCA1 p.Cys1477Arg The total amount of newly generated nascent HDL, the [3 H]cholesterol large/small particle ratio, and the size of the larger particles were all the lowest for the C1477R mutant, intermediate for the W509S variant, and the highest for the wild-type ABCA1 (Fig. 7). Login to comment 194 ABCA1 p.Trp590Ser ABCA1 p.Cys1477Arg BHK cells expressing wild-type ABCA1, ABCA1(W590S), or ABCA1(C1477R) were plated at the same density, labeled with [3 H]cholesterol, treated with 10 nM mifepristone, and exposed to 10 òe;g/ml of human apoAI to generate nascent HDL. Login to comment 195 ABCA1 p.Trp590Ser ABCA1 p.Cys1477Arg The decreased size of gel-filtration elution peaks indicates that W590S and especially C1477R mutations dramatically reduced the ability of ABCA1 to facilitate nascent HDL assembly. Login to comment