The photoaffinity probe 8-N3[alpha-32P]ATP labels the ATP-binding sites of rabbit neutrophil and skeletal muscle actin.
8-Azido-[alpha-32P]ATP (8-N3-ATP) was used as a photoaffinity label for ATP binding sites in the subcellular fractions of rabbit peritoneal neutrophils. The radioactive 8-N3-ATP was specifically incorporated into one major protein of 43 kDa. The isoelectric point, molecular mass and subcellular distribution of this labeled protein closely resemble those of the actin. 8-N3-[alpha-32P]ATP was further tested as a photoaffinity label for the ATP binding site in the purified rabbit skeletal muscle G-actin. The radioactive 8-N3-ATP was specifically incorporated into the actin band in SDS-polyacrylamide gel. The results indicate that 8-N3-ATP can be used as a photoaffinity label for actin.
D000199:Actins / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000378:metabolism*; D000818:Animals; D001386:Azides / Q000378:metabolism*; D001665:Binding Sites; D007526:Isoelectric Point; D009504:Neutrophils / Q000378:metabolism*; D010777:Photochemistry; D011817:Rabbits; D013347:Subcellular Fractions / Q000032:analysis
D000199:Actins; D001386:Azides; C017040:8-azidoadenosine 5'-triphosphate; D000255:Adenosine Triphosphate