ABCMdb

PMID: 11279083

Aleksandrov L, Mengos A, Chang X, Aleksandrov A, Riordan JR
Differential interactions of nucleotides at the two nucleotide binding domains of the cystic fibrosis transmembrane conductance regulator.
J Biol Chem. 2001 Apr 20;276(16):12918-23. Epub 2001 Jan 29., 2001-04-20 [PubMed]

Sentences

No. Mutations Sentence Comment

23 ABCC7 p.Lys1250Ala ABCC7 p.Lys464Ala EXPERIMENTAL PROCEDURES Materials- BHK-21 cells stably expressing wild-type human CFTR were prepared and maintained as described previously (20) as were cells expressing the K464A and K1250A mutants. Login to comment 63 ABCC7 p.Lys1250Ala ABCC7 p.Lys464Ala A, membranes from BHK cells expressing wild-type and the K1250A (15 ␮g of protein) and K464A variants (60 ␮g of protein) were incubated with 20 ␮M 8-azido-[␣-32 P]ATP in the presence of 0.5 mM orthovanadate and processed as under "Experimental Procedures." Login to comment 65 ABCC7 p.Lys464Ala Note that 4 times more K464A membrane protein was used than wild type and K1250. Login to comment 66 ABCC7 p.Lys464Ala The cells expressing K464A had also been grown at 26 °C to promote maturation of the mutant CFTR. B, 15 ␮g of BHK membrane protein containing wild-type CFTR was incubated at 0 °C for 1 h with 50 ␮g/ml purified immunoglobulin from each of the mouse monoclonal antibodies. Login to comment 69 ABCC7 p.Lys1250Ala (K1250A) is reduced only a small amount compared with wild type indicating that normally there may be little hydrolysis and trapping at NBD2. Login to comment 70 ABCC7 p.Lys464Ala In contrast the comparable substitution in NBD1 (K464A) greatly diminished labeling compared with wild type indicating that the majority of labeling and trapping may occur at NBD1. Login to comment