ABCMdb

ABCB3 p.Glu632Asp

None has been submitted yet.

Publications

PMID: 17068338 [PubMed] Perria CL et al: "Catalytic site modifications of TAP1 and TAP2 and their functional consequences."

No. Sentence Comment

73 The TAP2(E632Q/H661A)wasgeneratedusingTAP2(E632Q)in pPCR2.1 vector as the template. Login to comment
74 The primers used were H661A forward, 5Ј-CTGGTGATTGCTGCCAGGCTGCAGACA-3Ј and H661A reverse, 5Ј-TGTCTGCAGCCTGGCAGCAATCA- CCAG-3Ј. Login to comment
75 TAP2(E632D) and TAP2(H661Q) were generated using TAP2 in pPCR2.1 vector as the template. Login to comment
76 The primers used for TAP2(E632D) were E632D forward, 5Ј-CTCATCCTGGAT- GATGCTACTAGTGCC-3Ј and E632D reverse, 5Ј-GGCACTA- GTAGCATCATCCAGGATGAG-3Ј. Login to comment
77 The primers used for TAP2(H661Q) were H661Q forward, 5Ј-CTGGTGATTGCTC- AAAGGCTGCAGACA-3ЈandH661Qreverse,5Ј-TGTCTGCA- GCCTTTGAGCAATCACCAG-3Ј. Login to comment
78 TAP2(E632DH661Q) was generatedusingTAP2(E632D)asatemplateandusingtheprimers for TAP2(H661Q). Login to comment
79 Sequences encoding TAP2, TAP2(E632Q), TAP2(E632D), TAP2(H661Q), TAP2(E632D/H661Q), and TAP2(E632Q/ H662A) were excised from pPCR2.1 with BglII and XhoI and inserted into pMSCV 2.1 that was digested with BglII and XhoI. Login to comment
227 The abbreviations are: WT for wild type TAP2, EQ for TAP2(E632Q), EQHA for TAP2(E632Q/H661A), ED for TAP2(E632D), HQ for TAP2(H661Q), and EDHQ for TAP2(E632D/H661Q). Login to comment
241 The TAP2(E632D) and (H632Q) Mutations Result in Attenuated TAP Activity, whereas the TAP1(D668E/Q701H) Mutation Induces a Slight Increase in TAP Activity-In TAP1 sequences, Asp668 and Gln701 replace highly conserved glutamic acidandhistidineresidues,respectively,inotherABCtransporters (Fig. 1). Login to comment
244 To further examine this possibility, we analyzed the effects of TAP2(E632D), TAP2(H661Q), and TAP2(E632D/H661Q) mutants upon TAP activity in STF-1 cells. Login to comment
246 The TAP2(E632D) mutation (41% activity relative to wild type) affected TAP activity more significantly than the TAP2(H661Q) mutation (63% relative to wild type), and the double mutant (39% relative to wild type) exhibited a similar level of impairment as the TAP2(E632D) mutation. Login to comment
264 DISCUSSION Various modifications of TAP2 at Glu632 and His661 , including E632D and H661Q, significantly reduced the catalytic activity of TAP complexes (Figs. 2 and 4, A, C, E, and G). Login to comment
279 These data indicated a more important role for the switch region histidine in mediating ATP hydrolysis within isolated TAP1 NBD; however, at the TAP2 site of full-length TAP complexes, it appears that the TAP2(E632D) mutation causes a slightly greater loss in TAP activity compared with the TAP2(H661Q) mutation (Fig. 4E). Login to comment
290 Because the chimeric TAP1⅐T2MT1C complexes contained non-consensus residues (Asp668 and Gln701 ) at both nucleotide binding sites, the naturally occurring Glu Ͼ Asp and His Ͼ Gln alterations at these residues apparently did not completely abrogate the ability of the TAP1 site to hydrolyze ATP (consistent with the low but measurable activity of TAP2(E632D/H661Q); Fig. 4, E and G). Login to comment
72 The TAP2(E632Q/H661A)wasgeneratedusingTAP2(E632Q)in pPCR2.1 vector as the template. Login to comment
226 The abbreviations are: WT for wild type TAP2, EQ for TAP2(E632Q), EQHA for TAP2(E632Q/H661A), ED for TAP2(E632D), HQ for TAP2(H661Q), and EDHQ for TAP2(E632D/H661Q). Login to comment
240 The TAP2(E632D) and (H632Q) Mutations Result in Attenuated TAP Activity, whereas the TAP1(D668E/Q701H) Mutation Induces a Slight Increase in TAP Activity-In TAP1 sequences, Asp668 and Gln701 replace highly conserved glutamic acidandhistidineresidues,respectively,inotherABCtransporters (Fig. 1). Login to comment
243 To further examine this possibility, we analyzed the effects of TAP2(E632D), TAP2(H661Q), and TAP2(E632D/H661Q) mutants upon TAP activity in STF-1 cells. Login to comment
245 The TAP2(E632D) mutation (41% activity relative to wild type) affected TAP activity more significantly than the TAP2(H661Q) mutation (63% relative to wild type), and the double mutant (39% relative to wild type) exhibited a similar level of impairment as the TAP2(E632D) mutation. Login to comment
263 DISCUSSION Various modifications of TAP2 at Glu632 and His661 , including E632D and H661Q, significantly reduced the catalytic activity of TAP complexes (Figs. 2 and 4, A, C, E, and G). Login to comment
278 These data indicated a more important role for the switch region histidine in mediating ATP hydrolysis within isolated TAP1 NBD; however, at the TAP2 site of full-length TAP complexes, it appears that the TAP2(E632D) mutation causes a slightly greater loss in TAP activity compared with the TAP2(H661Q) mutation (Fig. 4E). Login to comment
289 Because the chimeric TAP1ዼT2MT1C complexes contained non-consensus residues (Asp668 and Gln701 ) at both nucleotide binding sites, the naturally occurring Glu b0e; Asp and His b0e; Gln alterations at these residues apparently did not completely abrogate the ability of the TAP1 site to hydrolyze ATP (consistent with the low but measurable activity of TAP2(E632D/H661Q); Fig. 4, E and G). Login to comment
224 The abbreviations are: WT for wild type TAP2, EQ for TAP2(E632Q), EQHA for TAP2(E632Q/H661A), ED for TAP2(E632D), HQ for TAP2(H661Q), and EDHQ for TAP2(E632D/H661Q). Login to comment
238 The TAP2(E632D) and (H632Q) Mutations Result in Attenuated TAP Activity, whereas the TAP1(D668E/Q701H) Mutation Induces a Slight Increase in TAP Activity-In TAP1 sequences, Asp668 and Gln701 replace highly conserved glutamic acidandhistidineresidues,respectively,inotherABCtransporters (Fig. 1). Login to comment
261 DISCUSSION Various modifications of TAP2 at Glu632 and His661 , including E632D and H661Q, significantly reduced the catalytic activity of TAP complexes (Figs. 2 and 4, A, C, E, and G). Login to comment
276 These data indicated a more important role for the switch region histidine in mediating ATP hydrolysis within isolated TAP1 NBD; however, at the TAP2 site of full-length TAP complexes, it appears that the TAP2(E632D) mutation causes a slightly greater loss in TAP activity compared with the TAP2(H661Q) mutation (Fig. 4E). Login to comment
287 Because the chimeric TAP1ዼT2MT1C complexes contained non-consensus residues (Asp668 and Gln701 ) at both nucleotide binding sites, the naturally occurring Glu b0e; Asp and His b0e; Gln alterations at these residues apparently did not completely abrogate the ability of the TAP1 site to hydrolyze ATP (consistent with the low but measurable activity of TAP2(E632D/H661Q); Fig. 4, E and G). Login to comment