Six-fold rotational symmetry of ClpQ, the E-coli homolog of the 20S proteasome, and its ATP-dependent activator, ClpY

被引:93
作者
Kessel, M
Wu, WF
Gottesman, S
Kocsis, E
Steven, AC
Maurizi, MR
机构
[1] NIAMSD,STRUCT BIOL LAB,BETHESDA,MD 20892
[2] NCI,NIH,CELL BIOL LAB,BETHESDA,MD 20892
[3] NCI,NIH,MOL BIOL LAB,BETHESDA,MD 20892
[4] HEBREW UNIV JERUSALEM,HADASSAH MED SCH,DEPT MEMBRANE & ULTRASTRUCT RES,IL-91010 JERUSALEM,ISRAEL
基金
美国国家卫生研究院;
关键词
Clp protease; HslU/HsiV; ATP-dependent protease; proteasome; rotational symmetry; image analysis;
D O I
10.1016/S0014-5793(96)01261-6
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
ClpQ (HslV) is a homolog of the beta-subunits of the 20S proteasome. In E. coli, it is expressed from an operon that also encodes ClpY (HsIU), an ATPase homologous to the protease chaperone, ClpX. ClpQ (subunit M(r) 19 000) and ClpY (subunit M(r) 49 000) were purified separately as oligomeric proteins with molecular weights of similar to 220 000 and similar to 350 000, respectively, estimated by gel filtration. Mixtures of ClpY and ClpQ displayed ATP-dependent proteolytic activity against casein, and a complex of the two proteins was isolated by gel filtration in the presence of ATP. Image processing of negatively stained electron micrographs revealed strong six-fold rotational symmetry for both ClpY and ClpQ, suggesting that the subunits of both proteins are arranged in hexagonal rings. The molecular weight of ClpQ combined with its symmetry is consistent with a double hexameric ring, whereas the data on ClpY suggest only one such ring. The symmetry mismatch previously observed between hexameric ClpA and heptameric ClpP in the related ClpAP protease is apparently not reproduced in the symmetry-matched ClpYQ system.
引用
收藏
页码:274 / 278
页数:5
相关论文
共 32 条
  • [1] An Escherichia coli chromosomal ''addiction module'' regulated by 3',5'-bispyrophosphate: A model for programmed bacterial cell death
    Aizenman, E
    EngelbergKulka, H
    Glaser, G
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1996, 93 (12) : 6059 - 6063
  • [2] THE CRYSTAL-STRUCTURE OF THE BACTERIAL CHAPERONIN GROEL AT 2.8-ANGSTROM
    BRAIG, K
    OTWINOWSKI, Z
    HEGDE, R
    BOISVERT, DC
    JOACHIMIAK, A
    HORWICH, AL
    SIGLER, PB
    [J]. NATURE, 1994, 371 (6498) : 578 - 586
  • [3] SEQUENCE-ANALYSIS OF 4 NEW HEAT-SHOCK GENES CONSTITUTING THE HSLTS IBPAB AND HSLVU OPERONS IN ESCHERICHIA-COLI
    CHUANG, SE
    BURLAND, V
    PLUNKETT, G
    DANIELS, DL
    BLATTNER, FR
    [J]. GENE, 1993, 134 (01) : 1 - 6
  • [4] SCANNING-TRANSMISSION ELECTRON-MICROSCOPY AND SMALL-ANGLE SCATTERING PROVIDE EVIDENCE THAT NATIVE ESCHERICHIA-COLI CLPP IS A TETRADECAMER WITH AN AXIAL PORE
    FLANAGAN, JM
    WALL, JS
    CAPEL, MS
    SCHNEIDER, DK
    SHANKLIN, J
    [J]. BIOCHEMISTRY, 1995, 34 (34) : 10910 - 10917
  • [5] THE MECHANISM AND FUNCTIONS OF ATP-DEPENDENT PROTEASES IN BACTERIAL AND ANIMAL-CELLS
    GOLDBERG, AL
    [J]. EUROPEAN JOURNAL OF BIOCHEMISTRY, 1992, 203 (1-2): : 9 - 23
  • [6] GOTTESMAN S, 1990, J BIOL CHEM, V265, P7886
  • [7] GOTTESMAN S, 1993, J BIOL CHEM, V268, P22618
  • [8] REGULATION BY PROTEOLYSIS - ENERGY-DEPENDENT PROTEASES AND THEIR TARGETS
    GOTTESMAN, S
    MAURIZI, MR
    [J]. MICROBIOLOGICAL REVIEWS, 1992, 56 (04) : 592 - 621
  • [9] THE 3-DIMENSIONAL STRUCTURE OF PROTEASOMES FROM THERMOPLASMA-ACIDOPHILUM AS DETERMINED BY ELECTRON-MICROSCOPY USING RANDOM CONICAL TILTING
    HEGERL, R
    PFEIFER, G
    PUHLER, G
    DAHLMANN, B
    BAUMEISTER, W
    [J]. FEBS LETTERS, 1991, 283 (01): : 117 - 121
  • [10] HOLM T, 1979, J MOL BIOL, V129, P359