Studies on the Mode of Action of Polyoxins |
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Authors: | Masahiro Hori Kazuo Kakiki Saburo Suzuki Tomomasa Misato |
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Institution: | 1. The Institute of Physical and Chemical Research, Wako-shi, Saitama Pref., Japan;2. Kaken Chemical Co., Ltd., Honkomagome, Bunkyo-ku, Tokyo, Japan |
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Abstract: | (1) Chitin-UDP acetylglucosaminyltransferase (E.C. 2.4.1.16., chitin synthetase) in the cell-free system from phytopathogenic fungus Piricularia oryzae, and effects of various polyoxins and related compounds on the enzyme activity were studied. Polyoxins A~M, polyoxin A derivatives, polyoxin C derivatives, 5′-amino-5′-deoxyuridine, uridine and thymidine inhibited equally the incorporation of N-acetylglucosamine (GlcNAc) from UDP-N-acetylglucosamine (UDP-GlcNAc) into chitin. (2) Competition between the above inhibitors and UDP-GlcNAc was observed by kinetic studies. The Km for UDP-GlcNAc was determined to be 3.3 × 10?3 m and the Ki values for polyoxins A~M, except polyoxin C, were found to be in the range of 3.3 × 10?5 m to 3.4 × 10?6 m. For polyoxin C, 5′-amino-5′-deoxyuridine and uridine, the Ki values of 2.7 × 10?3 m, 8.0 × 10?3 m and 3.0 × 10?3 m were given, respectively. The inhibitor constants for other related compounds were also calculated. (3) The values of binding affinity, ?ΔG, for formation of substrate- or inhibitor-enzyme complexes were calculated from the Km or Ki values. In addition, partial binding affinities, ?Δg, for certain moieties or groups of polyoxins were estimated from the ?ΔG. For instance, the ?ΔG values for UDP-GlcNAc and polyoxin L were 5.7 kcal/mole and 9.2 kcal/mole, respectively. And the ?Δg values for the nucleoside moiety (part I), the carbamylpolyoxamic acid moiety (part II) and the carboxyl group at C5′ position of polyoxin L were 5.2, 3.5 and 0.7 kcal/mole, respectively. (4) From the results obtained, the mechanism of action and relation between chemical structure and competitive inhibition of chitin synthetase were discussed. |
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Keywords: | hyperthermophilic aerobic archaeon Aeropyrum pernix K1 l-serine sulfhydrylase" target="_blank">O-phospho-l-serine sulfhydrylase stability to thermal stress and organic solvent electrostatic interaction |
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