Structure and Stability of Recombinant Protein Depend on the Extra N-terminal Methionine Residue: S6 Permutein from Direct and Fusion Expression Systems

Authors

Vladimir N. Uversky, Institute for Biological Instrumentation of the Russian Academy of SciencesFollow
Ziedulla Kh. Abdullaev, Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences
Alexander S. Arseniev, Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences
Eduard V. Bocharov, Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences
Dmitry A. Dolgikh, Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences
Ramil F. Latypov, Institute of Protein Research of the Russian Academy of Sciences
Tatiana N. Melnik, Institute of Protein Research of the Russian Academy of Sciences
Konstantin S. Vassilenko, Institute of Protein Researchof the Russian Academy of Sciences
Mikhail P. Kirpichnikov, Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences

Document Type

Article

Publication Date

1999

Keywords

Permuted Protein, Recombinant Protein, Protein Structure, Conformational Stability

Digital Object Identifier (DOI)

https://doi.org/10.1016/S0167-4838(99)00096-5

Abstract

Two permuted variants of S6 ribosomal protein were obtained in direct and fusion expression systems, respectively. The product of direct expression contained the extra N-terminal methionine residue. The structural properties and conformational stability of these permuteins were compared using 1-D 1H-NMR, circular dichroism, intrinsic fluorescence, differential scanning calorimetry and resistance to urea-induced unfolding. A pronounced difference in all the parameters studied has been demonstrated. This means that the structure of recombinant protein can be sensitive to peculiarities of the expression and purification procedures, leading particularly to the presence or absence of the Met at the first position in the target protein sequence.

Was this content written or created while at USF?

Yes

Citation / Publisher Attribution

Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology, v. 1432, issue 2, p. 324-332

Scholar Commons Citation

Uversky, Vladimir N.; Abdullaev, Ziedulla Kh.; Arseniev, Alexander S.; Bocharov, Eduard V.; Dolgikh, Dmitry A.; Latypov, Ramil F.; Melnik, Tatiana N.; Vassilenko, Konstantin S.; and Kirpichnikov, Mikhail P., "Structure and Stability of Recombinant Protein Depend on the Extra N-terminal Methionine Residue: S6 Permutein from Direct and Fusion Expression Systems" (1999). Molecular Medicine Faculty Publications. 651.
https://digitalcommons.usf.edu/mme_facpub/651