Ferguson Plot Analysis of Chaperone ClpB from Moderate Halophile

Teruo Akuta; Yui Tomioka; Tomoto Ura; Masataka Nakagawa; Tsutomu Arakawa

doi:10.1007/s10930-024-10245-6

Ferguson Plot Analysis of Chaperone ClpB from Moderate Halophile

Protein J. 2025 Jan 5. doi: 10.1007/s10930-024-10245-6. Online ahead of print.

Authors

Teruo Akuta¹, Yui Tomioka¹, Tomoto Ura^{2

3}, Masataka Nakagawa¹, Tsutomu Arakawa⁴

Affiliations

¹ Research and Development Division, Kyokuto Pharmaceutical Industrial Co., Ltd, 3333-26, Aza-Asayama, Kamitezuna Tahahagi, Ibaraki, 318-0004, Japan.
² Institute for Quantum Life Science, National Institutes for Quantum Science and Technology, 4-9-1 Anagawa, Inage-ku, Chiba, 263-8555, Japan.
³ Institute of Pure and Applied Sciences, University of Tsukuba, Tsukuba, Ibaraki, 305-8573, Japan.
⁴ Alliance Protein Laboratories, 13380 Pantera Road, San Diego, CA, 92130, USA. tarakawa2@aol.com.

PMID: 39755991
DOI: 10.1007/s10930-024-10245-6

Abstract

The Ferguson plot is a simple method for determining the molecular weight of native proteins and their complexes. In this study, we tested the validity of the Ferguson plot based on agarose native gel electrophoresis using multimeric chaperone protein, ClpB, derived from a moderate halophile that forms a native hexamer. The Ferguson plot showed a single band with a molecular weight of 1,500 kDa, approximately twice the size of the native hexamer. This result is consistent with the structure of other chaperons that form a double ring assembly comprising two hexameric units, i.e., a dodecamer. Supporting this, dynamic light scattering experiment showed two peaks, which likely correspond to the hexamer and dodecamer structures.

Keywords: Assembly; Chaperone; ClpB; Ferguson plot.