ABCG2 p.Ser187Ala
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PMID: 14645676
[PubMed]
Nakanishi T et al: "Functional characterization of human breast cancer resistance protein (BCRP, ABCG2) expressed in the oocytes of Xenopus laevis."
No.
Sentence
Comment
4
Injection of oocytes with cRNA containing mutations of serine 187 in the ATP-binding cassette signature motif (S187T or S187A) resulted in strong expression of the mutant forms; however, these oocytes were devoid of transporter activity.
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ABCG2 p.Ser187Ala 14645676:4:120
status: VERIFIED57 The PCR product was ligated into the pCR Blunt TOPO II TABLE 1 Summary of cDNA constructs made as templates for producing BCRP cRNA Construct Plasmid Features of the BCRP cRNA Produced RNA Polymerase Consensus Sequence Upstream from BCRP cDNA I pSP64poly(A) R482T-poly(A) SP6 II pCR Blunt TOPO II Kozak-R482T-poly(A) T7 III pSD64TR 5ЈUTR-Kozak-R482T-3ЈUTR-poly(A) other BCRP forms: 5ЈUTR-Kozak-R482-3ЈUTR-poly(A) 5ЈUTR-Kozak-R482T/S187T- 3ЈUTR-poly(A) 5ЈUTR-Kozak-R482T/ S187A-3ЈUTR-poly(A) SP6 III- pSD64TR 5ЈUTR-R482T-3ЈUTR-poly(A) SP6 5ЈUTR, 3ЈUTR, portions of the 5Ј- and 3Ј-UTR of the Xenopus laevis beta-globin gene; Kozak, modified sequences proximate to the start codon, as described under Materials and Methods; Poly(A), addition of a poly(A) tail, as described under Materials and Methods.
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ABCG2 p.Ser187Ala 14645676:57:513
status: VERIFIED60 Introduction of S187T and S187A Mutations into Human BCRP cDNA.
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ABCG2 p.Ser187Ala 14645676:60:26
status: VERIFIED64 The S187T primer pair was 5Ј-G TTT ATC CGT GGT GTG AC T GGA GGA GAA AG-3Ј and 5Ј-CT TTC TCC TCC AGT CAC ACC ACG GAT AAA C-3Ј, and the S187A primer pair was 5Ј-G TTT ATC CGT GGT GTG GC T GGA GGA GAA AG-3Ј and 5Ј-CT TTC TCC TCC AGC CAC ACC ACG GAT AAA C-3Ј.
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ABCG2 p.Ser187Ala 14645676:64:158
status: VERIFIED163 To determine whether dimerization is required for BCRP activity, a mutant construct was created by substituting the highly conserved serine at residue 187 in the ABC signature motif with threonine (R482T/S187T) or alanine (R482T/S187A).
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ABCG2 p.Ser187Ala 14645676:163:229
status: VERIFIED199 Expression of dominant-negative construct was confirmed by Western blot (A) and by DNR accumulation (25 M, B) in control or R482T-expressing oocytes (f), R482T/S187T- expressing oocytes (o), or R482T/S187A-expressing oocytes (gray o).
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ABCG2 p.Ser187Ala 14645676:199:208
status: VERIFIED233 Hence, we made two mutations of serine 187 in this signature motif of BCRP R482T, one to an amino acid that, like serine, has a polar side chain (threonine, S187T) and the other to a nonpolar side chain amino acid (alanine, S187A).
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ABCG2 p.Ser187Ala 14645676:233:224
status: VERIFIED241 Although it is possible that the inhibition caused by the S187A/T mutants was the result of the mutant saturating a limiting and essential oocyte cellular component crucial for protein transport or maturation, it is more likely that the inhibition was analogous to a "dominant-negative" effect of the S187T mutant.
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ABCG2 p.Ser187Ala 14645676:241:58
status: VERIFIED
PMID: 26136557
[PubMed]
Woodward OM et al: "ABCG2: the molecular mechanisms of urate secretion and gout."
No.
Sentence
Comment
35
Using a Xenopus oocyte expression system, we demonstrated ABCG2 to be a high-capacity urate transporter with ABCG2-mediated c-14 uric acid efflux highly dependent on the intracellular concentration, and could be blocked by a specific ABCG2 inhibitor, FTC, or with a single amino acid substitution, S187A.
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ABCG2 p.Ser187Ala 26136557:35:298
status: NEW