Evaluating the Role of Puckering and Fluorine Atom in Stability and Folding of Fluoroproline Containing Proteins

다국어 초록 (Multilingual Abstract)

In the past decade, numerous studies have been reported that the residue specific incorporation of fluorine containing analogs into protein can enhance the stability of protein. On the other hand, the incorporation of fluoroproline can enhance both stability and refolding rate of recombinant proteins. The objective of this study was to determine the reason behind the enhanced stability and refolding rate of protein by comparing GFP variants containing fluoroproline or hydroxyproline. The fluorine atom of 4-fluoroproline played a significant role in enhancing stability, and Cγ-endo puckering property of (4S)-4-fluoroproline and (4S)-4- hydroxyproline plays a key role in enhancing protein refolding rate.

참고문헌 (Reference)

1 Budisa, N., "Wiltschi, Residue-specific global fluorination of Candida antarctica lipase B in Pichia pastoris" 6 : 1630-1639, 2010

2 Ravikumar, Y., "Unnatural amino acid mutagenesis-based enzyme engineering" 33 : 462-470, 2015

3 Zheng T. Y., "Thermodynamic consequences of incorporating 4-substituted proline derivatives into a small helical protein" 49 : 4255-4263, 2010

4 Steiner, T., "Synthetic biology of proteins: tuning GFPs folding and stability with fluoroproline" 3 : e1680-, 2018

5 Balasubramanian, R., "Studies on the conformation of amino acids. VI. Conformation of the proline ring as observed in crystal structures of amino acids and peptides" 3 : 25-33, 1971

6 Holzberger, B., "Structural insights into the potential of 4-fluoroproline to modulate biophysical properties of proteins" 3 : 2924-2931, 2012

7 Hodges, J. A., "Stereoelectronic and steric effects in the collagen triple helix: Toward a code for strand association" 127 : 15923-15932, 2005

8 Shoulders, M. D., "Stereoelectronic and steric effects in side chains preorganize a protein main chain" 107 : 559-564, 2010

9 Holzberger, B., "Replacing 32 proline residues by a noncanonical amino acid results in a highly active DNA polymerase" 132 : 15708-15713, 2010

10 Link, A. J., "Reassignment of sense codons in vivo" 36 : 291-298, 2005

11 Budisa, N., "Prolegomena to future experimental efforts on genetic code engineering by expanding its amino acid repertoire" 43 : 6426-6463, 2004

12 Hoesl, M. G., "Lipase congeners designed by genetic code engineering" 3 : 213-221, 2011

13 Ravikumar, Y., "Incorporating unnatural amino acids to engineer biocatalysts for industrial bioprocess applications" 10 : 1862-1876, 2015

14 Renner, C., "Fluoroprolines as tools for protein design and engineering" 40 : 923-925, 2001

15 Odar, C., "Fluoro amino acids: A rarity in nature, yet a prospect for protein engineering" 10 : 427-446, 2015

16 Biava, H., "Evolution of fluorinated enzymes: An emerging trend for biocatalyst stabilization" 14 : 340-351, 2014

17 Doerfel, L. K., "Entropic contribution of elongation factor P to Pro line positioning at the catalytic center of the ribosome" 137 : 12997-13006, 2015

18 Edwardraja, S., "Enhancing the thermal stability of a single-chain Fvfragment by in vivo global fluorination of the proline residues" 7 : 258-265, 2011

19 Deepankumar, K., "Enhancing the biophysical properties of mRFP1 through incorporation of fluoroproline" 440 : 509-514, 2013

20 Nagasundarapandian, S., "Engineering protein sequence composition for folding robustness renders efficient noncanonical amino acid incorporations" 11 : 2521-2524, 2010

21 Larregola, M., "Congeneric bioadhesive mussel foot proteins designed by modified prolines revealed a chiral bias in unnatural translation" 421 : 646-650, 2012

22 Dietz, D., "Applying γ-substituted prolines in the foldon peptide: Polarity contradicts preorganization" 16 : 403-406, 2015