ABCMdb

PMID: 7518437

Chang XB, Hou YX, Jensen TJ, Riordan JR
Mapping of cystic fibrosis transmembrane conductance regulator membrane topology by glycosylation site insertion.
J Biol Chem. 1994 Jul 15;269(28):18572-5., [PubMed]

Sentences

No. Mutations Sentence Comment

22 ABCC7 p.Asn900Asp ABCC7 p.Asn900Asp ABCC7 p.Asn894Asp ABCC7 p.Asn894Asp A fragment inwild- type pNUT-CFTR (21, 22) was replaced by a PCR fragment (between HpaI site atnucleotide 2463 andDraIII site atnucleotide 3328)coding for the N894D and N900D changes to produce aCFTR with no consensus glycosylation sites on extracytoplasmic loops (designated ELO,Fig. Login to comment 23 ABCC7 p.Glu115Ser 1B).A PCR fragment coding for the E115S change was introduced into EL0 to generate a consensus glycosylation site on extracellular loop 1 (ELI, Fig.1B).Fourdifferent PCR fragmentscontaininginsertions coding for peptides NTS, GSNNSH, HRNQS, and NTS were inserted into EL0 at extracytoplasmic loops 2, 3, 5, and 6, respectively, to generate potential glycosylation sites on each putative extracellular loop (Fig. 1B). Login to comment 24 ABCC7 p.Leu953Asn ABCC7 p.Val1056Asn ABCC7 p.Met156Asn PCR fragments coding for the changes M156N, L953N, and V1056N were introduced into EL0 to generate potential glycosylation sites on cytoplasmic loop 1(CLI,Fig. Login to comment 64 ABCC7 p.Glu115Ser The extent of sequence perturbation employed to insert these sites rangedfrom a single aminoacid change (E115S)in EL1 to the insertionof 6 additional residues inEL3. Login to comment