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°C,...

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Bibliographic Details
Published in:Journal of biochemistry (Tokyo) 1989-10, Vol.106 (4), p.552-554
Main Authors: Yamamoto, Akira, Toda, Hiroko, Sakiyama, Fumio
Format: Article
Language:English
Subjects:
Amino Acids - analysis
Analytical, structural and metabolic biochemistry
Biological and medical sciences
chickens
Chromatography
Egg White - analysis
Fundamental and applied biological sciences. Psychology
General aspects, investigation methods
Hydrazines
lysozyme
Muramidase - analysis
Proteins
Proteins - analysis
Spectrophotometry, Ultraviolet
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Summary: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°C, the recovery of the carboxyl-terminal leueine 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 earboxyl-terminus of a protein is about 50pmol, as judged so far with hen egg-white lysozyme. The carboxyl-termini of several proteins were determined using this novel procedure.
ISSN:0021-924X
1756-2651
DOI:10.1093/oxfordjournals.jbchem.a122892