ABCMdb

PMID: 19297616

Oancea G, O'Mara ML, Bennett WF, Tieleman DP, Abele R, Tampe R
Structural arrangement of the transmission interface in the antigen ABC transport complex TAP.
Proc Natl Acad Sci U S A. 2009 Apr 7;106(14):5551-6. Epub 2009 Mar 18., [PubMed]

Sentences

No. Mutations Sentence Comment

57 ABCB3 p.Glu602Ala ABCB3 p.Glu602Cys ABCB3 p.Glu602Asp Remarkably, the X-loop mutations showed a reduced transport activity, with 50% transport activity for E602C and 20% for E602D or E602A (Fig. 2B). Login to comment 58 ABCB3 p.Glu602Ala ABCB3 p.Glu602Cys ABCB3 p.Glu602Asp ABCB3 p.Glu602Arg Complete disruption of peptide transport was observed for the E602R mutant. Login to comment 59 ABCB3 p.Glu602Arg Complete disruption of peptide transport was observed for the E602R mutant. Login to comment 63 ABCB3 p.Glu602Cys Altogether, 24 single-cysteine TAP1 mutants were coexpressed with TAP2(E602C) and yielded similar expression levels (Fig. 3A). Login to comment 64 ABCB3 p.Glu602Cys Altogether, 24 single-cysteine TAP1 mutants were coexpressed with TAP2(E602C) and yielded similar expression levels (Fig. 3A). Login to comment 66 ABCB2 p.Gln277Cys Remarkably, all mutations in coupling helix 1, except for Q277C, interfered with the transport activity. Login to comment 67 ABCB2 p.Gln277Cys Remarkably, all mutations in coupling helix 1, except for Q277C, interfered with the transport activity. Login to comment 89 ABCB3 p.Glu602Cys TAP cross-linking conditions using copper phenanthroline (CuPhe) were established for the A381/E602C complex (Fig. 4A and Fig. S2). Login to comment 90 ABCB3 p.Glu602Cys TAP cross-linking conditions using copper phenanthroline (CuPhe) were established for the A381/E602C complex (Fig. 4A and Fig. S2). Login to comment 91 ABCB3 p.Glu602Cys TAP cross-linking conditions using copper phenanthroline (CuPhe) were established for the A381/E602C complex (Fig. 4A and Fig. S2). Login to comment 100 ABCB3 p.Glu602Cys These findings demonstrate the specificity of the cross-linking between the X-loop (E602C) of TAP2 and CL1/2 of TAP1. Login to comment 101 ABCB3 p.Glu602Cys These findings demonstrate the specificity of the cross-linking between the X-loop (E602C) of TAP2 and CL1/2 of TAP1. Login to comment 102 ABCB3 p.Glu602Cys These findings demonstrate the specificity of the cross-linking between the X-loop (E602C) of TAP2 and CL1/2 of TAP1. Login to comment 121 ABCB3 p.Glu602Cys Specific peptide binding and transport of TAP1(Cys-less)/TAP2(E602C) was normalized to 100%. Login to comment 122 ABCB3 p.Glu602Cys Specific peptide binding and transport of TAP1(Cys-less)/TAP2(E602C) was normalized to 100%. Login to comment 124 ABCB3 p.Glu602Cys Specific peptide binding and transport of TAP1(Cys-less)/TAP2(E602C) was normalized to 100%. Login to comment 129 ABCB2 p.Gln277Cys ABCB3 p.Glu602Cys ABCB3 p.Glu602Cys We chose 2 pairs of mutations, Q277C/E602C and A381C/E602C, containing a single cysteine in each coupling helix of TAP1. Login to comment 130 ABCB2 p.Gln277Cys ABCB3 p.Glu602Cys ABCB3 p.Glu602Cys We chose 2 pairs of mutations, Q277C/E602C and A381C/E602C, containing a single cysteine in each coupling helix of TAP1. Login to comment 131 ABCB3 p.Glu602Cys Strikingly, cross-linking of the A381C/E602C complex impeded both peptide binding and transport, in contrast to the combination with Cys-less TAP1 (Fig. 5B). Login to comment 132 ABCB2 p.Gln277Cys ABCB3 p.Glu602Cys ABCB3 p.Glu602Cys ABCB3 p.Glu602Cys Surprisingly, cross-linking of Q277C/E602C arrested TAP in a translocation-incompetent state, in which peptide binding was unaffected (Fig. 5C). Login to comment 133 ABCB2 p.Gln277Cys ABCB3 p.Glu602Cys ABCB3 p.Glu602Cys The transport activity of Cys-less/E602C is only slightly reduced due to irreversible oxidation. Login to comment 134 ABCB3 p.Glu602Cys ABCB3 p.Glu602Cys The transport activity of Cys-less/E602C is only slightly reduced due to irreversible oxidation. Login to comment 135 ABCB2 p.Gln277Cys ABCB3 p.Glu602Cys Surprisingly, cross-linking of Q277C/E602C arrested TAP in a translocation-incompetent state, in which peptide binding was unaffected (Fig. 5C). Login to comment 136 ABCB3 p.Glu602Cys The transport activity of Cys-less/E602C is only slightly reduced due to irreversible oxidation. Login to comment 161 ABCB3 p.Glu602Cys (B and C) The cross-linking of the single-cysteine mutants of CL1 (B) and CL2 (C) in TAP1 with E602C of TAP2. Login to comment 163 ABCB3 p.Glu602Cys (B and C) The cross-linking of the single-cysteine mutants of CL1 (B) and CL2 (C) in TAP1 with E602C of TAP2. Login to comment 165 ABCB3 p.Glu602Cys (B and C) The cross-linking of the single-cysteine mutants of CL1 (B) and CL2 (C) in TAP1 with E602C of TAP2. Login to comment 201 ABCB3 p.Glu602Cys Membranes (500 ␮g of protein) containing variants of TAP1 in combination with TAP2(E602C) were incubated in the presence or absence of CuPhe for 1 min at 4 °C. Login to comment 203 ABCB3 p.Glu602Cys Membranes (500 òe;g of protein) containing variants of TAP1 in combination with TAP2(E602C) were incubated in the presence or absence of CuPhe for 1 min at 4 &#b0;C. Login to comment 205 ABCB3 p.Glu602Cys Membranes (500 òe;g of protein) containing variants of TAP1 in combination with TAP2(E602C) were incubated in the presence or absence of CuPhe for 1 min at 4 &#b0;C. Login to comment