ABCMdb

ABCB1 p.Ser23Cys

None has been submitted yet.

Publications

PMID: 15709779 [PubMed] Kaur P et al: "Biochemical characterization of domains in the membrane subunit DrrB that interact with the ABC subunit DrrA: identification of a conserved motif."

No. Sentence Comment

76 Primers used for making S23C mutation are shown here as an example: Single cysteine substitution mutants were created at amino acid positions 4, 15, 23, 35, 44, 53, 70, 80, 92, 107, 116, 129, 149, 160, 173, 213, 236, 249, 270, and 282 in DrrB. Login to comment
77 The mutants were named S4C, S15C, S23C, S35C, A44C, C260S up: 5'-GGCCTGGTCCTGTCCGTGTCGGCAGGG-3' C260S dn: 5'-CCCTGCCGACACGGACAGGACCAGGCC-3' S23C up: 5'- CGGACGGTGCTGTGCGCGGGTGAACGG-3' S23C dn: 5'- CCGTTCACCCGCGCACAGCACCGTCCG-3' V53C, T70C, S80C, V92C, S107C, V116C, A129C, T149C, A160C, V173C, S213C, S236C, T249C, A270C, and A282C, respectively. Login to comment
156 FIGURE 3: Chemical cross-linking between wild-type DrrA and DrrB containing cysteine substitution C260S or S23C. Login to comment
157 The cell membrane fraction containing the DrrA and DrrB (C260S or S23C) proteins was subjected to chemical cross-linking using different concentrations of GMBS, as described in Materials and Methods. Login to comment
163 (E) A reaction containing the cross-linked S23C sample was analyzed by SDS-PAGE, as described above. Login to comment
166 The lane containing the S23C sample was vertically spliced into halves. Each half was then probed with either the anti-DrrA or anti-DrrB antibodies. Detection was done by the chemiluminescence method. Login to comment
174 The N-terminal mutant S23C was the first mutant where a cross-linked product of DrrA and DrrB was identified (Figure 3). Login to comment
180 The 60 kDa cross-linked species did not appear in the cysteine-less (C260S) sample on addition of the cross-linker (Figure 3A,C) or in the S23C control sample where DMSO alone was added instead of the cross-linker (Figure 3B,D, lane 1). Login to comment
181 To further verify if the same cross-linked species was detected by both anti-DrrA and anti-DrrB antibodies, a lane containing the cross-linked S23C sample was spliced vertically into halves. Each half was probed with either the FIGURE 4: Chemical cross-linking between wild-type DrrA and DrrB containing cysteine substitutions in the N-terminal cytoplasmic tail, the transmembrane domains TM1 and TM2, and the cytoplasmic loop C1. Login to comment
193 The location of the cross-linked species is marked as A+B. (A) S23C (positive control), A129C, T149C, A160C. Login to comment
196 Table 1: Doxorubicin Resistance of E. coli N43 Cells Expressing Wild Type DrrA with DrrB Containing Different Cysteine Substitutionsa amt of dox, µg/mLdomain of DrrB location of cysteine 0 4 6 8 (wild type) C260 +++ +++ +++ ++ N-terminus S15C +++ +++ +++ ++ N-terminus S23C +++ +++ +++ ++ N-terminus S35C +++ +++ ++ ++ N-terminus A44C +++ +++ ++ + TM 1 V53C +++ ++ ++ - TM 1 T70C +++ + + + P 1 S80C +++ ++ + - TM 2 V92C +++ ( - - TM 2 S107C +++ +++ ++ ++ C 1 V116C +++ +++ +++ ++ TM 3 A129C +++ ++ + - TM 4 T149C +++ ++ ++ + C 2 A160C +++ +++ ++ + TM 5 V173C +++ +++ ++ + TM 6 S213C +++ +++ ++ + C 3 S236C +++ ++ + - TM 7 S249C +++ + + - TM 8 A270C +++ +++ ++ ++ C-terminus A282C +++ +++ ++ ++ cysteine-less DrrB C260S +++ +++ +++ +++ vector only pSU2718 ++++ ( - - a Legend: +++, very good growth; ++, good growth; +, some growth; -, no growth. Login to comment
205 It should be pointed out that the disappearance of the 43 kDa band happens in all the samples, irrespective of the presence (S23C) or absence (C260S) of a cysteine in DrrB (Figure 3A). Login to comment
206 Since the 60 kDa cross-linked species is not seen in C260S (even though the 43 kDa band disappears), it clearly indicates that the cross-linked species seen in S23C is not the result of cross-linking between DrrB and this 43 kDa unrelated protein. Login to comment
209 S23C, which showed a specific 60 kDa cross-linked species of DrrA and DrrB (described above), was used as a positive control in further cross-linking experiments. Login to comment
211 As seen with S23C, all the other cysteine substitution mutants tested in the N-terminal tail, S4C, S15C, S35C, and S44C, also showed the appearance of the 60 kDa species on cross-linking with GMBS by Western blot analysis (Figure 4A,B). Login to comment

PMID: 18086475 [PubMed] Alenius M et al: "Gene polymorphism influencing treatment response in psychotic patients in a naturalistic setting."

No. Sentence Comment

127 The second polymorphism studied was the change from serine to cysteine at residue 23 of the serotonin receptor 2C gene (HTR2C) located on the X-chromosome. Login to comment