Single-point substitution F97M leads to in cellulo crystallization of the biphotochromic protein moxSAASoti.

Autor: Marynich NK; A.N. Bach Institute of Biochemistry, Federal Research Centre 'Fundamentals of Biotechnology' of the Russian Academy of Sciences, Moscow, Russia., Boyko KM; A.N. Bach Institute of Biochemistry, Federal Research Centre 'Fundamentals of Biotechnology' of the Russian Academy of Sciences, Moscow, Russia., Matyuta IO; A.N. Bach Institute of Biochemistry, Federal Research Centre 'Fundamentals of Biotechnology' of the Russian Academy of Sciences, Moscow, Russia., Minyaev ME; N.D. Zelinsky Institute of Organic Chemistry Russian Academy of Sciences, Moscow, Russia., Khadiyatova AA; A.N. Bach Institute of Biochemistry, Federal Research Centre 'Fundamentals of Biotechnology' of the Russian Academy of Sciences, Moscow, Russia., Popov VO; A.N. Bach Institute of Biochemistry, Federal Research Centre 'Fundamentals of Biotechnology' of the Russian Academy of Sciences, Moscow, Russia., Savitsky AP; A.N. Bach Institute of Biochemistry, Federal Research Centre 'Fundamentals of Biotechnology' of the Russian Academy of Sciences, Moscow, Russia. Electronic address: apsavitsky@inbi.ras.ru.
Jazyk: angličtina
Zdroj: Biochemical and biophysical research communications [Biochem Biophys Res Commun] 2024 Nov 05; Vol. 732, pp. 150419. Date of Electronic Publication: 2024 Jul 17.
DOI: 10.1016/j.bbrc.2024.150419
Abstrakt: To enhance the photoconversion performance of biphotochromic moxSAASoti protein, a substitution F97 M was introduced. In addition to enhancing the target properties, this substitution also resulted in the crystallization of the recombinant protein within living HeLa cells (also referred to as in cellulo crystallization). The phenomenon of protein crystallization in living cells is not unique, yet the mechanisms and application of in cellulo crystallization remain significant for further research. However, in cellulo crystallization is atypical for fluorescent proteins and detrimental for their biotechnological application. The objective of this study was to elucidate the underlying mechanisms responsible for the crystallization of moxSAASoti F97M in cellulo. For this purpose, the crystal structure of the green form of biphotochromic protein moxSAASoti F97M was determined at high resolution, which surprisingly has a space group, different from those of parent mSAASoti C21N . The analysis provided allowed to propose a mechanism of new crystal contacts formation, which might be a cause of in cellulo protein crystallization.
Competing Interests: Declaration of competing interest The authors declare no conflicts of interest.
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Databáze: MEDLINE
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