차희정 (University of California-Berkeley)

한빛사논문

조회185

University of California-Berkeley

CV File 129 kb

Nano Lett., May 09 2024, | https://doi.org/10.1021/acs.nanolett.4c01043 STORM Super-Resolution Visualization of Self-Assembled γPFD Chaperone Ultrastructures in Methanocaldococcus jannaschii

Hee-Jeong Cha ^1#, Changdong He ^2#, Dominic J Glover ³, Ke Xu ^2,4*, Douglas S Clark ^1,4*

¹Department of Chemical and Biomolecular Engineering, University of California─Berkeley, Berkeley, California 94720, United States.

²Department of Chemistry, University of California─Berkeley, Berkeley, California 94720, United States.

³School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, NSW 2052, Australia.

⁴Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, 1 Cyclotron Road, Berkeley, California 94720, United States.

^#Co-first authors

^*Corresponding Authors : Ke Xu, Douglas S. Clark

Abstract

Gamma-prefoldin (γPFD), a unique chaperone found in the extremely thermophilic methanogen Methanocaldococcus jannaschii, self-assembles into filaments in vitro, which so far have been observed using transmission electron microscopy and cryo-electron microscopy. Utilizing three-dimensional stochastic optical reconstruction microscopy (3D-STORM), here we achieve ∼20 nm resolution by precisely locating individual fluorescent molecules, hence resolving γPFD ultrastructure both in vitro and in vivo. Through CF647 NHS ester labeling, we first demonstrate the accurate visualization of filaments and bundles with purified γPFD. Next, by implementing immunofluorescence labeling after creating a 3xFLAG-tagged γPFD strain, we successfully visualize γPFD in M. jannaschii cells. Through 3D-STORM and two-color STORM imaging with DNA, we show the widespread distribution of filamentous γPFD structures within the cell. These findings provide valuable insights into the structure and localization of γPFD, opening up possibilities for studying intriguing nanoscale components not only in archaea but also in other microorganisms.

논문정보

형식Research article
게재일2024년 05월 (BRIC 등록일 2024-05-27)
연구진국외 연구진
분야 생명과학 > 생물공학

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