ABCMdb

ABCB9 p.Lys545Met

Predicted by SNAP2:	A: D (95%), C: D (95%), D: D (95%), E: D (95%), F: D (95%), G: D (95%), H: D (95%), I: D (95%), L: D (95%), M: D (95%), N: D (95%), P: D (95%), Q: D (95%), R: D (91%), S: D (95%), T: D (95%), V: D (95%), W: D (95%), Y: D (95%),
Predicted by PROVEAN:	A: D, C: D, D: D, E: D, F: D, G: D, H: D, I: D, L: D, M: D, N: D, P: D, Q: D, R: D, S: D, T: D, V: D, W: D, Y: D,

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Publications
[hide] Examination of drug resistance activity of human T... Biochem Biophys Res Commun. 2006 May 5;343(2):597-601. Epub 2006 Mar 10. Ohashi-Kobayashi A, Ohashi K, Du WB, Omote H, Nakamoto R, Al-Shawi M, Maeda M
Examination of drug resistance activity of human TAP-like (ABCB9) expressed in yeast.
Biochem Biophys Res Commun. 2006 May 5;343(2):597-601. Epub 2006 Mar 10., [PMID:16554024]

Abstract [show]
A half-type ABC transporter, human TAP-like (hTAPL) tagged with histidine cluster, was expressed in budding yeast protease-deficient strain BJ5457, and the effect of expression for resistance to peptide compounds including antibiotics and proteinase inhibitor was examined. Among these compounds, the yeast expressing hTAPL exhibits high sensitivity to valinomycin, a monovalent cation ionophore. A mutation in Walker A motif, which lost ATP-binding activity of hTAPL, eliminated the enhanced sensitivity to valinomycin. These findings suggest that the transport activity of hTAPL is important for conferring high valinomycin-sensitive phenotype to yeast.

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Sentences [show]
No. Sentence Comment
23 For YEpTAPL(K546M)His, the DraIII-NgoMI fragment from phTAPL(K545M) plasmid, which has a mutation of Lys (AAG) at position 546 to Met (AtG) by PCR technique reported by Imai et al. [11] using mutated primers, was excised and replaced by the DraIII-NgoMI fragment of YEpTAPLHis. Login to comment
83 For this purpose, we constructed an expression plasmid for hTAPL(K545M) harboring mutation in the Walker A motif, and it was introduced into yeast. Login to comment
84 Prior to drug sensitivity assay, we determined the ATP-binding capabilities of hTAPL and hTAPL(K545M). Login to comment
85 Expression of hTAPL(K545M) was similar in amounts compared with that of hTAPL by means of Western blotting. Login to comment
87 3A and B, hTAPL bound ATP-agarose, whereas hTAPL(K545M) did not. Login to comment
94 TAPL p.c. P P S S - + - + (ATP) - + - + (ATP) TAPL K545M p.c. P P S S hTAPL(K545M) Raf61 hMDR hTAPL 5103 1103 2102 4101 8100 100µM valinomycin (cells/µl) A B Fig. 3. Login to comment
96 (A) ATP-binding activity was examined on hTAPL and hTAPL(K545M) (mutation in Walker A motif). Login to comment
97 Plasmid containing hTAPL or hTAPL(K545M) was transfected to yeast and the membrane fractions were prepared. Login to comment
101 (B) Sensitivity to valinomycin was examined on the yeast cells expressing hTAPL, hTAPL(K545M), or carrying control empty vector. Login to comment
103 Next, we examined valinomycin sensitivity of the yeast cell expressing hTAPL(K545M). Login to comment
82 For this purpose, we constructed an expression plasmid for hTAPL(K545M) harboring mutation in the Walker A motif, and it was introduced into yeast. Login to comment
86 3A and B, hTAPL bound ATP-agarose, whereas hTAPL(K545M) did not. Login to comment
93 TAPL p.c. P P S S - + - + (ATP) - + - + (ATP) TAPL K545M p.c. P P S S hTAPL(K545M) Raf61 hMDR hTAPL 5 10 3 1 10 3 2 10 2 4 10 1 8 10 0 100&#b5;M valinomycin (cells/&#b5;l) A B Fig. 3. Login to comment
95 (A) ATP-binding activity was examined on hTAPL and hTAPL(K545M) (mutation in Walker A motif). Login to comment
100 (B) Sensitivity to valinomycin was examined on the yeast cells expressing hTAPL, hTAPL(K545M), or carrying control empty vector. Login to comment
102 Next, we examined valinomycin sensitivity of the yeast cell expressing hTAPL(K545M). Login to comment