Identification of Essential Amino Acid Residues of an .ALPHA.-Amylase Inhibitor from Phaseolus vulgaris White Kidney Beans.
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概要
- 論文の詳細を見る
Kidney bean (Phaseolus vulgaris) α-amylase inhibitors, which are bivalent inhibitors with the subunit stoichiometry of (αβ)_??_ complex, have been inferred to contain unique arginine, tryptophan, and tyrosine residues essential for the inhibitory activity. To test the validity of this inference, an attempt was made to identify the essential amino acid residues of a white kidney bean (P. vulgaris) α-amylase inhibitor (PHA-I) by using the chemical modification technique combined with amino acid sequencing and mass spectrometry. Exhaustive modification of the arginine residues by phenylglyoxal did not lead to a marked loss of activity, suggesting that no arginine residue is directly associated with the inhibitory activity. N-Bromosuccinimide treatment of PHA-I in the presence or absence of a substrate α-amylase revealed the involvement of two tryptophan residues in α-amylase inhibition, and they were identified as Trp 188 of the β-subunit by amino acid sequencing and mass spectrometry of lysylendopeptidase peptides. Further, two tyrosine residues were preferentially modified either by N-acetylimidazole or by tetranitromethane, resulting in a concomitant loss of most of the PHA-I activity. Amino acid sequencing of the lysylendopeptidase peptides from a tetranitromethane-modified PHA-I identified Tyr 186 of the β-subunit as an essential residue.
- 社団法人 日本生化学会の論文
著者
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Nishimoto Tomoyuki
Department Of Agricultural Chemistry College Of Agriculture University Of Osaka Prefecture
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Nagai Kaoru
Department Of Applied Biological Chemistry College Of Agriculture Osaka Prefecture University
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Wada Yoshinao
Department Of Molecular Medicine Osaka Medical Center
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Wato Shin-ichi
Department Of Applied Biological Chemistry College Of Agriculture Osaka Pregecture University
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Hiramoto Sachiko
Department Of Applied Biological Chemistry College Of Agriculture Osaka Prefecture University
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Takahashi Takuya
Department Of Applied Biological Chemistry College Of Agriculture Osaka Prefecture University
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Yamaguchi Haruki
Department Of Agricultural Chemistry College Of Agriculture University Of Osaka Prefecture
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