Role of the nonheme Fe(II) center in the biosynthesis of the plant hormone ethylene

被引:98
作者
Rocklin, AM
Tierney, DL
Kofman, V
Brunhuber, NMW
Hoffman, BM
Christoffersen, RE
Reich, NO
Lipscomb, JD
Que, L
机构
[1] Univ Minnesota, Dept Chem & Biochem, Minneapolis, MN 55455 USA
[2] Univ Minnesota, Dept Mol Biol & Biophys, Minneapolis, MN 55455 USA
[3] Univ Minnesota, Ctr Met Biocatalysis, Minneapolis, MN 55455 USA
[4] Northwestern Univ, Dept Chem, Evanston, IL 60208 USA
[5] Univ Calif Santa Barbara, Dept Mol Cellular & Dev Biol, Santa Barbara, CA 93106 USA
[6] Univ Calif Santa Barbara, Dept Chem, Santa Barbara, CA 93106 USA
关键词
D O I
10.1073/pnas.96.14.7905
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
The final step of ethylene biosynthesis in plants is catalyzed by the enzyme 1-aminocyclopropane-1-carboxylic acid (ACC) oxidase (ACCO). In addition to ACC, Fe(II), O(2), CO(2), and ascorbate are required for in vitro enzyme activity. Direct evidence for the role of the Fe(II) center in the recombinant avocado ACCO has now been obtained through formation of enzyme (substrate or cofactor) NO complexes. These NO adducts convert the normally EPR-silent ACCO complexes into EPR-active species with structural properties similar to those of the corresponding O(2) complexes. It is shown here that the ternary Fe(II)ACCO . ACC . NO complex is readily formed, but no Fe(II)ACCO . ascorbate . NO complex could be observed, suggesting that ascorbate and NO are mutually exclusive in the active site. The binding modes of ACC and the structural analog alanine specifically labeled with (15)N or (17)O were examined by using Q-band electron nuclear double resonance (ENDOR). The data indicate that these molecules bind directly to the iron through both the alpha-amino and alpha-carboxylate groups. These observations are inconsistent with the currently favored mechanism for ACCO, in which it is proposed that both ascorbate and O(2) bind to the iron as a step in O(2) activation, We propose a different mechanism in which the iron serves instead to simultaneously bind ACC and O(2), thereby fixing their relative orientations and promoting electron transfer between them to initiate catalysis.
引用
收藏
页码:7905 / 7909
页数:5
相关论文
共 40 条
  • [21] Heterologous expression and site-directed mutagenesis of the 1-aminocyclopropane-1-carboxylate oxidase from kiwi fruit
    Lay, VJ
    Prescott, AG
    Thomas, PG
    John, P
    [J]. EUROPEAN JOURNAL OF BIOCHEMISTRY, 1996, 242 (02): : 228 - 234
  • [22] LIPSCOMB JD, 1992, MET IONS BIOL SYST, V28, P243
  • [23] LOWRY OH, 1951, J BIOL CHEM, V193, P265
  • [24] NUCLEOTIDE-SEQUENCE OF A RIPENING-RELATED CDNA FROM AVOCADO FRUIT
    MCGARVEY, DJ
    YU, H
    CHRISTOFFERSEN, RE
    [J]. PLANT MOLECULAR BIOLOGY, 1990, 15 (01) : 165 - 167
  • [25] NELSON MJ, 1987, J BIOL CHEM, V262, P12137
  • [26] Circular dichroism and magnetic circular dichroism spectroscopic studies of the non-heme ferrous active site in clavaminate synthase and its interaction with α-ketoglutarate cosubstrate
    Pavel, EG
    Zhou, J
    Busby, RW
    Gunsior, M
    Townsend, CA
    Solomon, EI
    [J]. JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1998, 120 (04) : 743 - 753
  • [27] PURIFICATION AND PROPERTIES OF THE APPLE FRUIT ETHYLENE-FORMING ENZYME
    PIRRUNG, MC
    KAISER, LM
    CHEN, J
    [J]. BIOCHEMISTRY, 1993, 32 (29) : 7445 - 7450
  • [28] Ethylene biosynthesis: processing of a substrate analog supports a radical mechanism for the ethylene-forming enzyme
    Pirrung, MC
    Cao, J
    Chen, JL
    [J]. CHEMISTRY & BIOLOGY, 1998, 5 (01): : 49 - 57
  • [29] A DILEMMA OF DIOXYGENASES (OR WHERE BIOCHEMISTRY AND MOLECULAR-BIOLOGY FAIL TO MEET)
    PRESCOTT, AG
    [J]. JOURNAL OF EXPERIMENTAL BOTANY, 1993, 44 (262) : 849 - 861
  • [30] Dioxygen activation by enzymes with mononuclear non-heme iron active sites
    Que, L
    Ho, RYN
    [J]. CHEMICAL REVIEWS, 1996, 96 (07) : 2607 - 2624