Multiple polypeptide forms observed in two-dimensional gels of Methylococcus capsulatus (Bath) polypeptides are generated during the separation procedure

被引:33
作者
Berven, FS
Karlsen, OA
Murrell, JC
Jensen, HB
机构
[1] Univ Bergen, Dept Mol Biol, N-5020 Bergen, Norway
[2] Univ Warwick, Dept Biol Sci, Coventry CV4 7AL, W Midlands, England
关键词
conforrhational equilibria; post-translational modifications; proteomics; trains of spots; two-dimensional gel electrophoresis;
D O I
10.1002/elps.200390091
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
We have examined two-dimensional electrophoresis (2-DE) gel maps of polypeptides from the Gram-negative bacterium Methylococcus capsulatus (Bath) and found the same widespread trains of spots as often reported in 2-DE gels of polypeptides of other Gram-negative bacteria. Some of the. trains of polypeptides, both from the outer membrane and soluble protein fraction, were shown to be generated during the separation procedure of 2-DE, and not by covalent post-translational modifications. The trains were found to be regenerated when rerunning individual polypeptide spots. The polypeptides analysed giving this type of trains were all found to be classified as stable polypeptides according to the instability index of Guruprasad et al. (Protein Eng. 1990, 4, 155-161). The phenomenon most likely reflects conformational equilibria of polypeptides arising from the experimental conditions used, and is a clear drawback of the standard 2-DE procedure,. making the gel picture unnecessarily complex to analyse.
引用
收藏
页码:757 / 761
页数:5
相关论文
共 36 条
  • [11] 2-C
  • [12] SIMPLIFIED METHOD FOR SILVER STAINING OF PROTEINS IN POLYACRYLAMIDE GELS AND THE MECHANISM OF SILVER STAINING
    HEUKESHOVEN, J
    DERNICK, R
    [J]. ELECTROPHORESIS, 1985, 6 (03) : 103 - 112
  • [13] Structure and function of bacterial outer membrane proteins: barrels in a nutshell
    Koebnik, R
    Locher, KP
    Van Gelder, P
    [J]. MOLECULAR MICROBIOLOGY, 2000, 37 (02) : 239 - 253
  • [14] CLEAVAGE OF STRUCTURAL PROTEINS DURING ASSEMBLY OF HEAD OF BACTERIOPHAGE-T4
    LAEMMLI, UK
    [J]. NATURE, 1970, 227 (5259) : 680 - +
  • [15] Lutter P, 2001, ELECTROPHORESIS, V22, P2888, DOI 10.1002/1522-2683(200108)22:14<2888::AID-ELPS2888>3.0.CO
  • [16] 2-C
  • [17] Proteomic analysis of the Escherichia coli outer membrane
    Molloy, MP
    Herbert, BR
    Slade, MB
    Rabilloud, T
    Nouwens, AS
    Williams, KL
    Gooley, AA
    [J]. EUROPEAN JOURNAL OF BIOCHEMISTRY, 2000, 267 (10): : 2871 - 2881
  • [18] Molloy MP, 2001, ELECTROPHORESIS, V22, P1686, DOI 10.1002/1522-2683(200105)22:9<1686::AID-ELPS1686>3.0.CO
  • [19] 2-L
  • [20] EFFECTS OF HEATING IN DODECYL-SULFATE SOLUTION ON CONFORMATION AND ELECTROPHORETIC MOBILITY OF ISOLATED MAJOR OUTER MEMBRANE PROTEINS FROM ESCHERICHIA-COLI K-12
    NAKAMURA, K
    MIZUSHIMA, S
    [J]. JOURNAL OF BIOCHEMISTRY, 1976, 80 (06) : 1411 - 1422