Crystallographic phases through genetic engineering: experiences with colicin A.
Colicins are antibiotic proteins that kill sensitive Escherichia coli cells. The structure of the pore-forming fragment of colicin A has been solved to 2.5 A resolution using the techniques of X-ray crystallography and genetic engineering. Site-directed mutagenesis was used to construct a number of cysteine-containing mutant proteins, one of which yielded an excellent mercurial derivative. Our experiences suggest strategies for obtaining useful heavy-atom derivatives for protein crystallography using genetic engineering techniques.
Tucker
A D
AD
European Molecular Biology Laboratory, Heidelberg, FRG.
Baty
D
D
Parker
M W
MW
Pattus
F
F
Lazdunski
C
C
Tsernoglou
D
D
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000032:analysis / Q000302:isolation & purification*; D001483:Base Sequence; D003087:Colicins; D003461:Crystallography; D004926:Escherichia coli; D008958:Models, Molecular; D008969:Molecular Sequence Data; D015202:Protein Engineering
D001426:Bacterial Proteins; D003087:Colicins; C007104:colicin immunity proteins