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Investigation of the interaction between enzyme and inhibitor by the combination of chemical modification, electrospray ionization mass spectrometry and frit‐fast atom bombardment liquid chromatography/mass spectrometry
Author(s) -
Akashi Satoko,
Niitsu Uno,
Yuji Reiko,
Ide Hiroyuki,
Hirayama Kazuo
Publication year - 1993
Publication title -
biological mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 121
eISSN - 1096-9888
pISSN - 1052-9306
DOI - 10.1002/bms.1200220205
Subject(s) - chemistry , fast atom bombardment , mass spectrometry , electrospray ionization , chemical modification , chromatography , reagent , chemical ionization , sample preparation in mass spectrometry , chemical structure , organic chemistry , ionization , biochemistry , ion
The interaction between enzyme and its inhibitor, hen egg‐white lysozyme and tri‐ N ‐acetylglucosamine (NAG 3 ), was studied by the combination of chemical modification, enzymatic digestion, electrospray ionization mass spectrometry and frit–fast atom bombardment liquid chromatography/mass spectrometry. Chemical modification of amino groups, carboxyl groups, and indole groups was carried out independently. In the absence of NAG 3 , the carboxyl group in Asp 101 was modified by glycinamidation, and the indole group in Trp 62 was modified by Koshland reagent. In the presence of NAG 3 , the degree of modification of Asp 101 and Trp 62 decreased. It is suggested that Asp 101 and Trp 62 are involved in the interaction with NAG 3 . The result is consistent with the one obtained by x‐ray crystallography. It is indicated that the combination of chemical modification and mass spectrometry may be effective for the investigation of the binding reaction of enzyme to inhibitor and of protein–protein interaction.