Analysis of F factor TraD membrane topology by use of gene fusions and trypsin-sensitive insertions.
This report describes a procedure for characterizing membrane protein topology which combines the analysis of reporter protein hybrids and trypsin-sensitive 31-amino-acid insertions generated by using transposons ISphoA/in and ISlacZ/in. Studies of the F factor TraD protein imply that the protein takes on a structure with two membrane-spanning sequences and amino and carboxyl termini facing the cytoplasm. It was possible to assign the subcellular location of one region for which the behavior of fused reporter proteins was ambiguous, based on the trypsin cleavage behavior of a 31-residue insertion.
Lee
M H
MH
Departments of Genetics, University of Washington, Seattle, Washington 98195-7360, USA.
Kosuk
N
N
Bailey
J
J
Traxler
B
B
Manoil
C
C
D000469:Alkaline Phosphatase / Q000032:analysis / Q000235:genetics; D003227:Conjugation, Genetic; D029968:Escherichia coli Proteins; D005144:F Factor; D017930:Genes, Reporter; D005816:Genetic Complementation Test; D008565:Membrane Proteins / Q000737:chemistry* / Q000235:genetics; D008958:Models, Molecular; D016254:Mutagenesis, Insertional; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins; D014357:Trypsin
D029968:Escherichia coli Proteins; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; C064957:traD protein, E coli; D000469:Alkaline Phosphatase; D014357:Trypsin