Modification of tryptophan residues of plant class III chitinases involved in enzyme activity

Masatsune Ishiguro, Takeshi Yamagami, Miho Tanigawa, Kazuki Tsutsumi, Gunki Funatsu, Takayuki Ohnuma, Yoichi Aso

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Abstract

Chemical modification of tulip bulb chitinase-1 (TBC-1) and pokeweed leaf chitinase (PLC-B) with N-bromosuccinimide (NBS) suggested the involvement of tryptophan residue (s) in the activity. In the case of TBC-1, at pH 4.0, about 1 mol of tryptophan residues out of 4 mol was oxidized with 5 mol of NBS per mol of TBC-1 and all 4 mol of tryptophan residues were oxidized with 15mol of NBS. At pH 4.5, about 3 mol of tryptophan were finally oxidized although the oxidation rate of the first tryptophan residue was same as at pH 4.0. In both cases, the oxidation of 1 mol of tryptophan residues caused 85% activity loss. Analysis of the oxytryptophan-containing peptides afforded to identify that Trp172 and Trp255 were those which reacted first with NBS at pH 4.0. When PLC-B was reacted with 16-folds molar of NBS at pH 4.0, all seven tryptophan residues were oxidized with a concomitant decrease of chitinase activity. Oxidized tryptophan residues were also determined as those at positions of 165 and 256 in PLC-B. From these results, it was inferred that the rapidly oxidized tryptophan residues in plant class HI chhitinases were involved in their chitinase activity.

Original languageEnglish
Pages (from-to)243-250
Number of pages8
JournalJournal of the Faculty of Agriculture, Kyushu University
Volume46
Issue number1
Publication statusPublished - 2001

Fingerprint

plant residues
chitinase
Tryptophan
tryptophan
Chitinases
Bromosuccinimide
enzyme activity
Enzymes
Tulipa
bulbs
Phytolacca americana
5-hydroxytryptophan
oxidation
class III chitinase
5-Hydroxytryptophan
peptides
Peptides

All Science Journal Classification (ASJC) codes

  • Agricultural and Biological Sciences (miscellaneous)

Cite this

Modification of tryptophan residues of plant class III chitinases involved in enzyme activity. / Ishiguro, Masatsune; Yamagami, Takeshi; Tanigawa, Miho; Tsutsumi, Kazuki; Funatsu, Gunki; Ohnuma, Takayuki; Aso, Yoichi.

In: Journal of the Faculty of Agriculture, Kyushu University, Vol. 46, No. 1, 2001, p. 243-250.

Research output: Contribution to journalArticle

Ishiguro, M, Yamagami, T, Tanigawa, M, Tsutsumi, K, Funatsu, G, Ohnuma, T & Aso, Y 2001, 'Modification of tryptophan residues of plant class III chitinases involved in enzyme activity', Journal of the Faculty of Agriculture, Kyushu University, vol. 46, no. 1, pp. 243-250.
Ishiguro, Masatsune ; Yamagami, Takeshi ; Tanigawa, Miho ; Tsutsumi, Kazuki ; Funatsu, Gunki ; Ohnuma, Takayuki ; Aso, Yoichi. / Modification of tryptophan residues of plant class III chitinases involved in enzyme activity. In: Journal of the Faculty of Agriculture, Kyushu University. 2001 ; Vol. 46, No. 1. pp. 243-250.
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