Molecular characterization of the 4′-phosphopantothenoylcysteine decarboxylase domain of bacterial Dfp flavoproteins

被引:28
作者
Kupke, T [1 ]
机构
[1] Univ Tubingen, Lehrstuhl Mikrobielle Genet, D-72076 Tubingen, Germany
关键词
D O I
10.1074/jbc.M103342200
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
The NH2-terminal domain of the bacterial flavoprotein Dfp catalyzes the decarboxylation of (R)-4'-phospho-N-pantothenoylcysteine to 4'-phosphopantetheine, a key step in coenzyme A biosynthesis, Dfp proteins, LanD proteins (for example EpiD, which is involved in epidermin biosynthesis), and the salt tolerance protein AtHAL3a from Arabidopsis thaliana are homooligomeric flavin-containing Cys decarboxylases (HFCD protein family), The crystal structure of the peptidyl-cysteine decarboxylase EpiD complexed with a pentapeptide substrate has recently been determined. The peptide is bound by an NH2-terminal substrate binding helix, residue Asn(117), which contacts the cysteine residue of the substrate, and a COOH-terminal substrate recognition clamp, The conserved motif G-G/S-I-A-X-Y-K of the Dfp proteins aligns partly with the substrate binding helix of EpiD. Point mutations within this motif resulted in loss of coenzyme binding (G14S) or in significant decrease of Dfp activity (G15A, I16L, A17D, K20N, K20Q), Exchange of Asn(125) of Dfp, which corresponds to Asn(117) of EpiD, and exchange of Cys(158), which is within the proposed substrate recognition clamp of Dfp, led to inactivity of the enzyme, Molecular analysis of the conditional lethality of the Escherichia coli dfp-707 mutant revealed that the single point mutation G11D of Dfp is related to decreased amounts of soluble Dfp protein at 37 degreesC.
引用
收藏
页码:27597 / 27604
页数:8
相关论文
共 20 条
  • [1] The X-ray structure of the FMN-binding protein AtHal3 provides the structural basis for the activity of a regulatory subunit involved in signal transduction
    Albert, A
    Martínez-Ripoll, M
    Espinosa-Ruiz, A
    Yenush, L
    Culiáñez-Macià, FA
    Serrano, R
    [J]. STRUCTURE, 2000, 8 (09) : 961 - 969
  • [2] Begley TP, 2001, VITAM HORM, V61, P157
  • [3] Crystal structure of the peptidyl-cysteine decarboxylase EpiD complexed with a pentapeptide substrate
    Blaesse, M
    Kupke, T
    Huber, R
    Steinbacher, S
    [J]. EMBO JOURNAL, 2000, 19 (23) : 6299 - 6310
  • [4] Arabidopsis thaliana AtHAL3:: a flavoprotein related to salt and osmotic tolerance and plant growth
    Espinosa-Ruiz, A
    Bellés, JM
    Serrano, R
    Culiáñez-Macià, FA
    [J]. PLANT JOURNAL, 1999, 20 (05) : 529 - 539
  • [5] Purification and characterization of phosphopantetheine adenylyltransferase from Escherichia coli
    Geerlof, A
    Lewendon, A
    Shaw, WV
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (38) : 27105 - 27111
  • [6] JACKOWSKI S, 1996, ESCHERICHIA COLI SAL, V1, P687
  • [7] Molecular characterization of lantibiotic-synthesizing enzyme EpiD reveals a function for bacterial Dfp proteins in coenzyme A biosynthesis
    Kupke, T
    Uebele, M
    Schmid, D
    Jung, G
    Blaesse, M
    Steinbacher, S
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 2000, 275 (41) : 31838 - 31846
  • [8] PURIFICATION AND CHARACTERIZATION OF EPID, A FLAVOPROTEIN INVOLVED IN THE BIOSYNTHESIS OF THE LANTIBIOTIC EPIDERMIN
    KUPKE, T
    STEVANOVIC, S
    SAHL, HG
    GOTZ, F
    [J]. JOURNAL OF BACTERIOLOGY, 1992, 174 (16) : 5354 - 5361
  • [9] KUPKE T, 1994, J BIOL CHEM, V269, P5653
  • [10] Arabidopsis thaliana flavoprotein AtHAL3a catalyzes the decarboxylation of 4′-phosphopantothenoylcysteine to 4′-phosphopantetheine, a key step in coenzyme A biosynthesis
    Kupke, T
    Hernández-Acosta, P
    Steinbacher, S
    Culiáñez-Macià, FA
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 2001, 276 (22) : 19190 - 19196