Preparation and properties of an affinity support for purification of cyclic AMP receptor protein from Escherichia coli.
Reaction of cyclic AMP with 1,1'-carbonyldiimidazole produces an intermediate which reacts with primary amines to provide a stable 2'-O-carbamyl derivative. This chemistry has been used to tether cyclic AMP to a Sepharose gel. The resulting affinity support has been used to effect a simple, nondenaturing purification of cyclic AMP receptor protein from crude cell extracts.
Corrie
J E
JE
National Institute for Medical Research, London, United Kingdom.
Pizza
C
C
Makwana
J
J
King
R W
RW
D000345:Affinity Labels; D001426:Bacterial Proteins / Q000302:isolation & purification*; D002352:Carrier Proteins / Q000302:isolation & purification*; D002846:Chromatography, Affinity / Q000379:methods; D000242:Cyclic AMP / Q000031:analogs & derivatives; D004926:Escherichia coli / Q000737:chemistry*; D007093:Imidazoles; D011953:Receptors, Cyclic AMP / Q000302:isolation & purification*; D012685:Sepharose; D014157:Transcription Factors / Q000187:drug effects / Q000302:isolation & purification* / Q000378:metabolism
D000345:Affinity Labels; D001426:Bacterial Proteins; D002352:Carrier Proteins; D007093:Imidazoles; D011953:Receptors, Cyclic AMP; D014157:Transcription Factors; C006900:N,N-carbonyldiimidazole; D012685:Sepharose; D000242:Cyclic AMP