Vapor-phase hydrazinolysis for microdetermination of carboxyl-terminal amino acids of proteins.
The "vapor-phase" hydrazinolysis method was devised for the microdetermination of the carboxyl-terminal residue of a protein. With this method, a polypeptide sample is degraded with vaporized hydrazine. The optimum conditions for hen egg-white lysozyme were established to be 2 to 4 h at 90 or 100 degrees C, the recovery of the carboxyl-terminal leucine being about 70%. With this vapor-phase method, side reactions are reduced and the time of hydrazinolysis is shortened. The limit of quantitation for the carboxyl-terminus of a protein is about 50 pmol, as judged so far with hen egg-white lysozyme. The carboxyl-termini of several proteins were determined using this novel procedure.
Yamamoto
A
A
Institute for Protein Research, Osaka University.
Toda
H
H
Sakiyama
F
F
D000596:Amino Acids / Q000032:analysis*; D002845:Chromatography; D004529:Egg White / Q000032:analysis; D006834:Hydrazines; D009113:Muramidase / Q000032:analysis; D011506:Proteins / Q000032:analysis*; D013056:Spectrophotometry, Ultraviolet
D000596:Amino Acids; D006834:Hydrazines; D011506:Proteins; D009113:Muramidase