ABCMdb

PMID: 16487063

Qian F, Wei D, Liu J, Yang S
Molecular model and ATPase activity of carboxyl-terminal nucleotide binding domain from human P-glycoprotein.
Biochemistry (Mosc). 2006;71 Suppl 1:S18-24, 1-2., [PubMed]

Sentences

No. Mutations Sentence Comment

12 ABCB1 p.Gln1118Ala The functional role of Gln1118 was investigated by the studying the biochem ical properties of recombinant NBD2 and its mutant Q1118A. Login to comment 45 ABCB1 p.Gln1118Ala Construction of the expression plasmid encoding NBD2 and its mutant Q1118A. Login to comment 51 ABCB1 p.Gln1118Ala For site directed muta genesis to introduce a Q1118A mutation, nucleotide mutations at positions 3352 (C→G) and 3353 (A→C) were achieved by sequence overlap PCR methodology. Login to comment 54 ABCB1 p.Gln1118Ala The sequence of plasmid containing NBD2 Q1118A mutant was confirmed by sequencing. Login to comment 55 ABCB1 p.Gln1118Ala Expression, renaturation, and purification of NBD2 and its mutant Q1118A. Login to comment 56 ABCB1 p.Gln1118Ala Escherichia coli BL21 cells con taining NBD2 and its mutant Q1118A were cultured in 200 ml of 2YTA medium (16 g/liter tryptone, 10 g/liter yeast extract, 5 g/liter NaCl, and 100 µg/ml ampicillin) at 37°C with vigorous shaking until the absorbance at 600 nm reached 1.2 units. Login to comment 124 ABCB1 p.Gln1118Ala Expression and purification of NBD2 and its mutant Q1118A. Login to comment 127 ABCB1 p.Gln1118Ala The Q1118A mutation was introduced into the NBD2 sequence by site directed mutagenesis. Login to comment 128 ABCB1 p.Gln1118Ala Recombinant proteins encoded by pGEX4T 1 NBD2 and its mutant Q1118A based plas mids were expressed in E. coli cells. Login to comment 129 ABCB1 p.Gln1118Ala Expression and purification of the NBD2 and its mutant Q1118A are illustrated in Fig. 6. Login to comment 135 ABCB1 p.Gln1118Ala NBD2 protein and mutant Q1118A protein yielded equal purities and in similar amounts. Login to comment 140 ABCB1 p.Gln1118Ala Expression and purification of the NBD2 and its mutant Q1118A. Login to comment 142 ABCB1 p.Gln1118Ala Expression of mutant Q1118A protein before (lane ) and after (lane 4) IPTG induction. Login to comment 143 ABCB1 p.Gln1118Ala Purified NBD2 protein (lane 5) and its mutant Q1118A protein (lane 6). Login to comment 144 ABCB1 p.Gln1118Ala S22 QIAN et al. BIOCHEMISTRY (Moscow) Vol. 71 Suppl. 1 2006 ATPase activity of NBD2 and its mutant Q1118A. Login to comment 148 ABCB1 p.Gln1118Ala The ATPase activities of the Q1118A mutant and that of the wild type NBD2 were compared. Login to comment 165 ABCB1 p.Gln1118Ala The Q1118A mutant was affected in its ATPase activity but this residue might not participate the binding of ATP because the mutation had no influence on the affinity for ATP. Login to comment 174 ABCB4 p.Gln1118Ala a) ATP dependence of the ATPase activities of the isolated NBD2 (1) and its mutant Q1118A (2). Login to comment 176 ABCB4 p.Gln1118Ala b) Lineweaver-Burk plot of dependence of ATPase activity of NBD2 (1) and its mutant Q1118A (2). Login to comment