MOLECULAR-WEIGHTS OF GLYCOSYLATED AND NONGLYCOSYLATED FORMS OF RECOMBINANT HUMAN STEM-CELL FACTOR DETERMINED BY LOW-ANGLE LASER-LIGHT SCATTERING

被引:40
作者
ARAKAWA, T [1 ]
LANGLEY, KE [1 ]
KAMEYAMA, K [1 ]
TAKAGI, T [1 ]
机构
[1] INST PROT RES, SUITA, OSAKA, JAPAN
关键词
D O I
10.1016/0003-2697(92)90042-6
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The molecular weight of recombinant human stem cell factor (SCF) was determined using a low-angle laser light scattering combined with a differential refractometer and a uv detector. The protein samples were applied to these detectors through a gel filtration column by a high-performance liquid chromatographic pump. The Chinese hamster ovary (CHO) cell-derived SCF gave a molecular weight of 53,000 for the entire molecule and 35,000 for the protein moiety only at pH 7.0, indicating that the CHO cell-derived protein is glycosylated by 34%. Since the molecular weight of the polypeptide is 18,600, the results demonstrate that the CHO cell-derived SCF forms a dimer. The molecular weight of Escherichia coli-derived SCF was determined to be 39,000, similar to the above value (35,000). Essentially identical molecular weights were obtained at pH 3.0, indicating no dissociation of the dimer. © 1992.
引用
收藏
页码:53 / 57
页数:5
相关论文
共 28 条
  • [1] MOLECULAR-CLONING OF MAST-CELL GROWTH-FACTOR, A HEMATOPOIETIN THAT IS ACTIVE IN BOTH MEMBRANE-BOUND AND SOLUBLE FORMS
    ANDERSON, DM
    LYMAN, SD
    BAIRD, A
    WIGNALL, JM
    EISENMAN, J
    RAUCH, C
    MARCH, CJ
    BOSWELL, HS
    GIMPEL, SD
    COSMAN, D
    WILLIAMS, DE
    [J]. CELL, 1990, 63 (01) : 235 - 243
  • [2] ANDREWS RG, 1991, BLOOD, V78, P1975
  • [3] ARAKAWA T, 1991, J BIOL CHEM, V266, P18942
  • [4] ACTIVATION OF THE HUMAN C-KIT PRODUCT BY LIGAND-INDUCED DIMERIZATION MEDIATES CIRCULAR ACTIN REORGANIZATION AND CHEMOTAXIS
    BLUMEJENSEN, P
    CLAESSONWELSH, L
    SIEGBAHN, A
    ZSEBO, KM
    WESTERMARK, B
    HELDIN, CH
    [J]. EMBO JOURNAL, 1991, 10 (13) : 4121 - 4128
  • [5] THE PROTO-ONCOGENE C-KIT ENCODING A TRANSMEMBRANE TYROSINE KINASE RECEPTOR MAPS TO THE MOUSE W-LOCUS
    CHABOT, B
    STEPHENSON, DA
    CHAPMAN, VM
    BESMER, P
    BERNSTEIN, A
    [J]. NATURE, 1988, 335 (6185) : 88 - 89
  • [6] MAST-CELL GROWTH-FACTOR MAPS NEAR THE STEEL LOCUS ON MOUSE CHROMOSOME-10 AND IS DELETED IN A NUMBER OF STEEL ALLELES
    COPELAND, NG
    GILBERT, DJ
    CHO, BC
    DONOVAN, PJ
    JENKINS, NA
    COSMAN, D
    ANDERSON, D
    LYMAN, SD
    WILLIAMS, DE
    [J]. CELL, 1990, 63 (01) : 175 - 183
  • [7] DAS SK, 1982, J BIOL CHEM, V257, P13679
  • [9] TRANSMEMBRANE FORM OF THE KIT LIGAND GROWTH-FACTOR IS DETERMINED BY ALTERNATIVE SPLICING AND IS MISSING IN THE SI(D) MUTANT
    FLANAGAN, JG
    CHAN, DC
    LEDER, P
    [J]. CELL, 1991, 64 (05) : 1025 - 1035
  • [10] THE DOMINANT-WHITE SPOTTING (W) LOCUS OF THE MOUSE ENCODES THE C-KIT PROTO-ONCOGENE
    GEISSLER, EN
    RYAN, MA
    HOUSMAN, DE
    [J]. CELL, 1988, 55 (01) : 185 - 192