B01-3：Structural Characterization of Protein Complexes by Native Mass Spectrometry in Integrated Bio-metal Science
Satoko Akashi
（Graduate School of Medical Life Science, Yokohama City University）
Lab website
To understand the function of metal-bound proteins, structural characterization, including mass determination of the intact molecule in the native state, is significant. However, it is not easy to detect intact ions of the metal-bound proteins by mass spectrometry, without dissociation of metals that are essential for the function. In the present study, followings will be examined using native mass spectrometry (native MS) to establish the research basis of Bio-metal Science. 1) Development of a native MS method that enables definitive and accurate mass determination of intact metal-bound protein complexes with >50 kDa, leading to identification of the metals essential for function 2) Development of a screening system of specific ligands to a metal-bound protein using native MS under crude biological conditions 3) Characterization of conformational changes of a protein upon metal binding using ion mobility mass spectrometry 4) Native MS of membrane proteins containing metals With the native MS method mentioned above, collaboration within the IBmS area will be facilitated.
Major publications
K. Saikusa, A. Osakabe, D. Kato, S. Fuchigami, A. Nagadoi, Y. Nishimura, H. Kurumizaka, *S. Akashi
“Structural Diversity of Nucleosomes Characterized by Native Mass Spectrometry”
Anal. Chem., 2018, 90, 8217–8226
doi: 10.1021/acs.analchem.8b01649
D. Kato, A. Osakabe, Y. Arimura, Y. Mizukami, N. Horikoshi, K. Saikusa, S. Akashi, Y. Nishimura, S.-Y. Park, J. Nogami, K. Maehara, Y. Ohkawa, A. Matsumoto, H. Kono, R. Inoue, M. Sugiyama, *H. Kurumizaka
“Crystal structure of the overlapping dinucleosome composed of hexasome and octasome”
Science, 2017, 356, 205-208
doi: 10.1126/science.aak9867
S. Akashi and *K. M. Downard
“Effect of Charge on the Conformation of Highly Basic Peptides including the Tail Regions of Histone Proteins by Ion Mobility Mass Spectrometry”
Anal. Bioanal. Chem., 2016, 408, 6637-6648
doi: 10.1007/s00216-016-9777-4
Selected as a cover-page article
K. Saikusa, A. Nagadoi, K. Hara, S. Fuchigami, H. Kurumizaka, Y. Nishimura, *S. Akashi
“Mass spectrometric approach for characterizing the disordered tail regions of the histone H2A/H2B dimer”
Anal. Chem., 2015, 87, 2220-2227
doi: 10.1021/ac503689w
N. Azegami, K. Saikusa, Y. Todokoro, H. Kurumizaka, A. Nagadoi, *Y. Nishimura, *S. Akashi
“Conclusive evidence of the reconstituted hexasome proved by native mass spectrometry”
Biochemistry, 2013, 52, 5155-5157
doi: 10.1021/bi4005655
Selected as a highlighted article