The identification of apoptosis-related residues in human thymosin β-10 by mutational analysis and computational modeling

Seung Bae Rho, Keun Woo Lee, Taehoon Chun, Seung Hoon Lee, Kyoungsook Park, Je Ho Lee

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Thymosin β-10 (TB10) is an actin monomer-sequestering peptide that consists of 43 amino acid residues and that tends to form α-helical structures. Previously, we showed that the overexpression of TB10 dramatically increases the frequency of apoptosis in human ovarian cancer cells. To identify the critical residues responsible for TB10-mediated apoptosis, we used a series of computational methods. First, a three-dimensional structure of human TB10 was constructed using the homology modeling method with the calf thymosin β-9 NMR structure as a template. Although the sequences of both of these structures are almost identical, 200-ps molecular dynamics simulation results showed that their secondary structures differ. Analyses of molecular dynamics snapshot structures suggested that the TB10 structure is conformationally more complicated than the TB9 structure. The conserved 17LKKTET 22 motif region of TB10 was tested by Ala and Ser scanning mutagenesis using computational and biochemical methods, and 12 mutants were transfected into cancer cell lines and tested for their effects on growth arrest. Of the 12 mutants examined, only the Thr20 to Ser 20 mutation showed reduced growth arrest. These results strongly suggest that Thr20 is specifically required for actin sequestration by TB10 in ovarian cancer cells. These results may provide useful information for the development of a new ovarian cancer therapy.

Original languageEnglish
Pages (from-to)34003-34007
Number of pages5
JournalJournal of Biological Chemistry
Volume280
Issue number40
DOIs
Publication statusPublished - 2005 Oct 7
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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