Visualization of intrinsically disordered regions of proteins by high-speed atomic force microscopy

Atsushi Miyagi, Yasuo Tsunaka, Takayuki Uchihashi, Kouta Mayanagi, Susumu Hirose, Kosuke Morikawa, Toshio Ando

Research output: Contribution to journalArticle

71 Citations (Scopus)

Abstract

Intrinsically disordered (ID) regions of proteins are recognized to be involved in biological processes such as transcription, translation, and cellular signal transduction. Despite the important roles of ID regions, effective methods to observe these thin and flexible structures directly were not available. Herein, we use high-speed atomic force microscopy (AFM) to observe the heterodimeric FACT (facilitates chromatin transcription) protein, which is predicted to have large ID regions in each subunit. Successive AFM images of FACT on a mica surface, captured at rates of 5-17 frames per second, clearly reveal two distinct tail-like segments that protrude from the main body of FACT and fluctuate in position. Using deletion mutants of FACT, we identify these tail segments as the two major ID regions predicted from the amino acid sequences. Their mechanical properties estimated from the AFM images suggest that they have more relaxed structures than random coils. These observations demonstrate that this state-of-the-art microscopy method can be used to characterize unstructured protein segments that are difficult to visualize with other experimental techniques.

Original languageEnglish
Pages (from-to)1859-1866
Number of pages8
JournalChemPhysChem
Volume9
Issue number13
DOIs
Publication statusPublished - Sep 15 2008
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Atomic and Molecular Physics, and Optics
  • Physical and Theoretical Chemistry

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