Recombinant full-length murine prion protein, mPrP(23-231): purification and spectroscopic characterization

被引:152
作者
Hornemann, S
Korth, C
Oesch, B
Riek, R
Wider, G
Wuthrich, K
Glockshuber, R
机构
[1] ETH HONGGERBERG,INST MOL BIOL & BIOPHYS,CH-8093 ZURICH,SWITZERLAND
[2] UNIV ZURICH,PRIONICS AG,CH-8057 ZURICH,SWITZERLAND
关键词
transmissible spongiform encephalopathies (TSEs); cellular prion protein; protein conformation; circular dichroism spectroscopy;
D O I
10.1016/S0014-5793(97)00921-6
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
The cellular prion protein of the mouse, mPrP(C), consists of 208 amino acids (residues 23-231), It contains a carboxy-terminal domain, mPrP(121-231), which represents an autonomous folding unit with three alpha-helices and a two-stranded antiparallel beta-sheet, We expressed the complete amino acid sequence of the prion protein, mPrP(23-231), in the cytoplasm of Escherichia coli, mPrP(23-231) was solubilized from inclusion bodies by 8 M urea, oxidatively refolded and purified to homogeneity by conventional chromatographic techniques, Comparison of near-UV circular dichroism, fluorescence and one-dimensional H-1-NMR spectra of mPrP(23-231) and mPrP(121-231) shows that the amino-terminal segment 23-120, which includes the five characteristic octapeptide repeats, does not contribute measurably to the manifestation of three-dimensional structure as detected by these techniques, indicating that the residues 121-231 might be the only polypeptide segment of PrPC with a defined three-dimensional structure. (C) 1997 Federation of European Biochemical Societies.
引用
收藏
页码:277 / 281
页数:5
相关论文
共 27 条
  • [1] DOES AGENT OF SCRAPIE REPLICATE WITHOUT NUCLEIC ACID
    ALPER, T
    CRAMP, WA
    HAIG, DA
    CLARKE, MC
    [J]. NATURE, 1967, 214 (5090) : 764 - &
  • [2] SECONDARY STRUCTURE-ANALYSIS OF THE SCRAPIE-ASSOCIATED PROTEIN PRP 27-30 IN WATER BY INFRARED-SPECTROSCOPY
    CAUGHEY, BW
    DONG, A
    BHAT, KS
    ERNST, D
    HAYES, SF
    CAUGHEY, WS
    [J]. BIOCHEMISTRY, 1991, 30 (31) : 7672 - 7680
  • [3] TISSUE SULFHYDRYL GROUPS
    ELLMAN, GL
    [J]. ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1959, 82 (01) : 70 - 77
  • [4] Prion protein (PrP) with amino-proximal deletions restoring susceptibility of PrP knockout mice to scrapie
    Fischer, M
    Rulicke, T
    Raeber, A
    Sailer, A
    Moser, M
    Oesch, B
    Brandner, S
    Aguzzi, A
    Weissmann, C
    [J]. EMBO JOURNAL, 1996, 15 (06) : 1255 - 1264
  • [5] CALCULATION OF PROTEIN EXTINCTION COEFFICIENTS FROM AMINO-ACID SEQUENCE DATA
    GILL, SC
    VONHIPPEL, PH
    [J]. ANALYTICAL BIOCHEMISTRY, 1989, 182 (02) : 319 - 326
  • [6] SELF-REPLICATION AND SCRAPIE
    GRIFFITH, JS
    [J]. NATURE, 1967, 215 (5105) : 1043 - &
  • [7] HORNEMANN S, 1996, J MOL BIOL, V262, P614
  • [8] Deadly conformations - Protein misfolding in prion disease
    Horwich, AL
    Weissman, JS
    [J]. CELL, 1997, 89 (04) : 499 - 510
  • [9] PROPOSED 3-DIMENSIONAL STRUCTURE FOR THE CELLULAR PRION PROTEIN
    HUANG, ZW
    GABRIEL, JM
    BALDWIN, MA
    FLETTERICK, RJ
    PRUSINER, SB
    COHEN, FE
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1994, 91 (15) : 7139 - 7143
  • [10] JARETT JT, 1993, CELL, V73, P1055