Phosphoamino acid analysis.
It is often valuable to identify the phosphorylated residue in a protein. In the case of proteins phosphorylated at serine, threonine, or tyrosine, this is readily accomplished by partial acid hydrolysis in HCl followed by two-dimensional thin-layer electrophoresis of the labeled phosphoamino acid, as described here. Phosphothreonine andphosphotyrosine are more stable to hydrolysis in alkali than are RNA andpho sphoserine. Therefore, a protocol for mild alkaline hydrolysis of protein samples is also provided to enhance the detection of phosphothreonine and phosphotyrosine. Although this procedure can be carried out with a protein eluted from a preparative gel and concentrated by trichloroacetic acid or acetone precipitation, it is most easily accomplished by transfer of the protein of interest to a PVDF membrane.
Sefton
B M
BM
The Salk Institute, San Diego, California, USA.
D000468:Alkalies / Q000737:chemistry; D004586:Electrophoresis; D006868:Hydrolysis; D019606:Phosphoamino Acids / Q000032:analysis*; D011145:Polyvinyls; D011506:Proteins / Q000737:chemistry; D005465:Fluorocarbon Polymers
D000468:Alkalies; D019606:Phosphoamino Acids; D011145:Polyvinyls; D011506:Proteins; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers