Identification of the glycosylation site of the adenovirus type 5 fiber protein

被引:16
作者
Cauet, G
Strub, JM
Leize, E
Wagner, E
Van Dorsselaer, A
Lusky, M
机构
[1] Transgene SA, F-67082 Strasbourg, France
[2] ECPM, Lab Spectrometrie Masse Bioorgan, F-67087 Strasbourg, France
关键词
D O I
10.1021/bi047702b
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 fiber protein purified from the pool of nonincorporated viral protein after infection of cells with adenovirus type 5 exists as two forms separable by reverse-phase HPLC. As determined by mass spectrometry, this heterogeneity results from a difference in one O-linked N-acetylglucosamine (GlcNac). A western blot analysis using a monoclonal antibody directed against the GlcNac motif showed that only one of the two forms reacted with the antibody, suggesting that one form carries a single GlcNac and the other form has none. The ratio of glycosylated to nonglycosylated forms of fiber, which is about 1, is conserved in assembled viruses. After digestion of glycosylated fiber with endoproteinase GluC, isolation of the glycosylated peptide by reverse-phase HPLC, and chemical derivatization using dimethylamine, the site of glycosylation was located in the fiber shaft at serine 109 by mass spectrometry. Elimination of glycosylation by site-directed mutagenesis of fiber should help to understand the function of this postranslational modification.
引用
收藏
页码:5453 / 5460
页数:8
相关论文
共 33 条
  • [1] Alving K, 1999, J MASS SPECTROM, V34, P395, DOI 10.1002/(SICI)1096-9888(199904)34:4<395::AID-JMS771>3.0.CO
  • [2] 2-1
  • [3] Isolation of a common receptor for coxsackie B viruses and adenoviruses 2 and 5
    Bergelson, JM
    Cunningham, JA
    Droguett, G
    KurtJones, EA
    Krithivas, A
    Hong, JS
    Horwitz, MS
    Crowell, RL
    Finberg, RW
    [J]. SCIENCE, 1997, 275 (5304) : 1320 - 1323
  • [4] LARGE-SCALE PREPARATION OF SOLUBLE ADENOVIRUS HEXON, PENTON AND FIBER ANTIGENS IN HIGHLY PURIFIED FORM
    BOULANGE.PA
    PUVION, F
    [J]. EUROPEAN JOURNAL OF BIOCHEMISTRY, 1973, 39 (01): : 37 - 42
  • [5] THE USE OF ADENOVIRAL VECTORS FOR GENE-THERAPY AND GENE-TRANSFER IN-VIVO
    BRAMSON, JL
    GRAHAM, FL
    GAULDIE, J
    [J]. CURRENT OPINION IN BIOTECHNOLOGY, 1995, 6 (05) : 590 - 595
  • [6] O-LINKED GLCNAC IN SEROTYPE-2 ADENOVIRUS FIBER
    CAILLETBOUDIN, ML
    STRECKER, G
    MICHALSKI, JC
    [J]. EUROPEAN JOURNAL OF BIOCHEMISTRY, 1989, 184 (01): : 205 - 211
  • [7] CHENG CS, 1982, J VIROL, V42, P932
  • [8] O-glycosylation of nuclear and cytosolic proteins -: Dynamic interplay between O-GlcNAc and O-phosphate
    Comer, FI
    Hart, GW
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 2000, 275 (38) : 29179 - 29182
  • [9] Identification of substituted sites on MUC5AC mucin motif peptides after enzymatic O-glycosylation combining β-elimination and fixed-charge derivatization
    Czeszak, X
    Ricart, G
    Tetaert, D
    Michalski, JC
    Lemoine, J
    [J]. RAPID COMMUNICATIONS IN MASS SPECTROMETRY, 2002, 16 (01) : 27 - 34
  • [10] Gahéry-Ségard H, 1998, J VIROL, V72, P2388