Selected article for: "density centrifugation and gradient density centrifugation"
Author: Qiao, Hui; Pelletier, Sandra L.; Hoffman, Lucas; Hacker, Jill; Armstrong, R. Todd; White, Judith M.
Title: Specific Single or Double Proline Substitutions in the “Spring-loaded” Coiled-Coil Region of the Influenza Hemagglutinin Impair or Abolish Membrane Fusion Activity
  • Document date: 1998_6_15
    • ID: 78fjem8s_38
    Snippet: We next asked whether the single Pro-substituted HAs, the mutants that were considered most likely to be impaired in fusion, form trimers at the cell surface. Cells expressing V55P, S71P, or L80P HA0 were treated with trypsin to cleave HA0. Cell lysates were prepared and subjected to sucrose density gradient centrifugation analysis. As seen in Fig. 3 A , all of the single Pro-substituted HAs comigrated with wt-HA in fractions 6-8 of the gradient,.....
    Document: We next asked whether the single Pro-substituted HAs, the mutants that were considered most likely to be impaired in fusion, form trimers at the cell surface. Cells expressing V55P, S71P, or L80P HA0 were treated with trypsin to cleave HA0. Cell lysates were prepared and subjected to sucrose density gradient centrifugation analysis. As seen in Fig. 3 A , all of the single Pro-substituted HAs comigrated with wt-HA in fractions 6-8 of the gradient, suggesting that they formed 9S trimers that were stable to sucrose density centrifugation in the presence of a nonionic detergent. All of the Ala-substituted mutants behaved the same.

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