EVIDENCE FOR AN ESSENTIAL HISTIDINE RESIDUE IN 4S-LIMONENE SYNTHASE AND OTHER TERPENE CYCLASES

被引:41
作者
RAJAONARIVONY, JIM
GERSHENZON, J
MIYAZAKI, J
CROTEAU, R
机构
[1] WASHINGTON STATE UNIV,INST BIOL CHEM,PULLMAN,WA 99164
[2] WASHINGTON STATE UNIV,GRAD PROGRAM PLANT PHYSIOL,PULLMAN,WA 99164
关键词
D O I
10.1016/0003-9861(92)90246-S
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
(4S)-Limonene synthase, isolated from glandular trichome secretory cell preparations of Mentha × piperita (peppermint) leaves, catalyzes the metal ion-dependent cyclization of geranyl pyrophosphate, via 3S-linalyl pyrophosphate, to (-)-(4S)-limonene as the principal product. Treatment of this terpene cyclase with the histidine-directed reagent diethyl pyrocarbonate at a concentration of 0.25 mm resulted in 50% loss of enzyme activity, and this activity could be completely restored by treatment of the preparation with 5 mm hydroxylamine. Inhibition with diethyl pyrocarbonate was distinguished from inhibition with thiol-directed reagents by protection studies with histidine and cysteine carried out at varying pH. Inactivation of the cyclase by dye-sensitized photooxidation in the presence of rose bengal gave further indication of the presence of a readily modified histidine residue. Protection of the enzyme against inhibition with diethyl pyrocarbonate was afforded by the substrate geranyl pyrophosphate in the presence of Mn2+, and by the sulfonium ion analog of the linalyl carbocation intermediate of the reaction in the presence of inorganic pyrophosphate plus Mn2+, suggesting that an essential histidine residue is located at or near the active site. Similar studies on the inhibition of other monoterpene and sesquiterpene cyclases with diethyl pyrocarbonate suggest that a histidine residue (or residues) may play an important role in catalysis by this class of enzymes. © 1992.
引用
收藏
页码:77 / 82
页数:6
相关论文
共 41 条
[1]   PURIFICATION AND CHARACTERIZATION OF THE MONOTERPENE CYCLASE GAMMA-TERPINENE SYNTHASE FROM THYMUS-VULGARIS [J].
ALONSO, WR ;
CROTEAU, R .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1991, 286 (02) :511-517
[2]  
ALONSO WR, 1992, J BIOL CHEM, V267, P7582
[3]   DIETHYL PYROCARBONATE - EXAMINATION OF ITS PROPERTIES IN BUFFERED SOLUTIONS WITH A NEW ASSAY TECHNIQUE [J].
BERGER, SL .
ANALYTICAL BIOCHEMISTRY, 1975, 67 (02) :428-437
[4]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[5]   ENZYMATIC FORMATION OF SESQUITERPENES [J].
CANE, DE .
CHEMICAL REVIEWS, 1990, 90 (07) :1089-1103
[6]  
CHAYET I, 1985, BIOORG CHEM, V12, P329
[7]   BIOSYNTHESIS AND CATABOLISM OF MONOTERPENOIDS [J].
CROTEAU, R .
CHEMICAL REVIEWS, 1987, 87 (05) :929-954
[8]   BIOSYNTHESIS OF MONOTERPENES - ENZYMATIC CONVERSION OF NERYL PYROPHOSPHATE TO 1,8-CINEOLE, ALPHA-TERPINEOL, AND CYCLIC MONOTERPENE HYDROCARBONS BY A CELL-FREE PREPARATION FROM SAGE (SALVIA-OFFICINALIS) [J].
CROTEAU, R ;
KARP, F .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1976, 176 (02) :734-746
[9]   MONOTERPENE BIOSYNTHESIS - MECHANISTIC EVALUATION OF THE GERANYL PYROPHOSPHATE-(-)-ENDO-FENCHOL CYCLASE FROM FENNEL (FOENICULUM-VULGARE) [J].
CROTEAU, R ;
MIYAZAKI, JH ;
WHEELER, CJ .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1989, 269 (02) :507-516