Engineering a beta-turn in green fluorescent protein to a foreign loop

Bharat Madan; Jayaraman Thangappan; Sun-Gu Lee

Journal of Industrial and Engineering Chemistry, Vol.33, 330-335, January, 2016

DOI10.1016/j.jiec.2015.10.027 Export Citation

Engineering a beta-turn in green fluorescent protein to a foreign loop

Bharat Madan, Jayaraman Thangappan, and Sun-Gu Lee^†

Department of Chemical Engineering, Pusan National University, Busan 609-735, South Korea

E-mail:

A type I beta-turn in green fluorescent protein (GFP) was engineered to a foreign loop. Molecular dynamics simulation study showed that the addition of foreign loop into GFP did not have a negative influence on the conformation stability of GFP structure, but the GFP variant with the foreign loop sequence was completely misfolded in real folding conditions. The co-incorporation of the enhancing mutations for GFP folding made it possible to generate a foldable and active GFP variant with the foreign loop sequence, although the folding efficiency and specific activity of the GFP were negatively affected by the introduced loop.

Keywords:Protein engineering;Turn and loop;Folding and misfolding;Green fluorescent protein

[References]

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Hu X, Wang H, Ke H, Kuhlman B, Proc. Natl. Acad. Sci. U. S. A., 104, 17668 (2007)
Yang F, Moss LG, Phillips GN, Nat. Biotechnol., 14, 1246 (1996)
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Cormack BP, Valdivia RH, Falkow S, Gene, 173, 33 (1996)
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Crameri A, Whitehorn EA, Tate E, Stemmer WP, Nat. Biotechnol., 14, 315 (1996)
Ito Y, Suzuki M, Husimi Y, Biochem. Biophys. Res. Commun., 264, 556 (1999)
Teerawanichpan P, Hoffman T, Ashe P, Datla R, Selvaraj G, Biochim. Biophys. Acta, 1770, 1360 (2007)
Patterson GH, Knobel SM, Sharif WD, Kain SR, Piston DW, Biophys. J., 73, 2782 (1997)
Chalfie M, Tu Y, Euskirchen G, Ward WW, Prasher DC, Science, 263(5148), 802 (1994)

[Referenced By]

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[1] Martinez JC, Pisabarro MT, Serrano L, Nat. Struct. Biol., 5, 721 (1998)

[2] McCallister EL, Alm E, Baker D, Nat. Struct. Biol., 7, 669 (2000)

[3] Sanchez IE, Protein Sci., 17, 183 (2008)

[4] Ohage EC, Graml W, Walter MM, Steinbacher S, Steipe B, Protein Sci., 6, 233 (1997)

[5] Mulligan-Pullyblank P, Spitzer JS, Gilden BM, Fetrow JS, J. Biol. Chem., 271, 8633 (1996)

[6] Palma R, Curmi PM, Protein Sci., 8, 913 (1999)

[7] Doss CGP, Rajith B, Garwasis N, Mathew PR, Raju AS, Apoorva K, William D, Sadhana NR, Himani T, Dike IP, Appl. Transl. Genomics, 1, 37 (2012)

[8] Nagasundarapandian S, Merkel L, Budisa N, Govindan R, Ayyadurai N, Sriram S, Yun H, Lee SG, Chembiochem, 11, 2521 (2010)

[9] Raghunathan G, Soundrarajan N, Sokalingam S, Yun H, Lee SG, PLOS ONE, 7, e51510 (2012)

[10] Raghunathan G, Sokalingam S, Soundrarajan N, Madan B, Munussami G, Lee SG, Mol. Biosyst., 9, 2379 (2013)

[11] Madan B, Sokalingam S, Raghunathan G, Lee SG, Proteins, 82, 2812 (2014)

[12] Hu X, Wang H, Ke H, Kuhlman B, Proc. Natl. Acad. Sci. U. S. A., 104, 17668 (2007)

[13] Yang F, Moss LG, Phillips GN, Nat. Biotechnol., 14, 1246 (1996)

[14] Pavoor TV, Cho YK, Shusta EV, Proc. Natl. Acad. Sci. U. S. A., 106, 11895 (2009)

[15] Cormack BP, Valdivia RH, Falkow S, Gene, 173, 33 (1996)

[16] Borgo B, Havranek JJ, Proc. Natl. Acad. Sci. U. S. A., 109, 1494 (2012)

[17] Bloom JD, Glassman MJ, PLoS Comput. Biol., 5, e10003 (2009)

[18] Worth CL, Preissner R, Blundell TL, Nucleic Acids Res., 39, W215 (2011)

[19] Pedelacq JD, Cabantous S, Tran T, Terwilliger TC, Waldo GS, Nat. Biotechnol., 24, 79 (2006)

[20] Crameri A, Whitehorn EA, Tate E, Stemmer WP, Nat. Biotechnol., 14, 315 (1996)

[21] Ito Y, Suzuki M, Husimi Y, Biochem. Biophys. Res. Commun., 264, 556 (1999)

[22] Teerawanichpan P, Hoffman T, Ashe P, Datla R, Selvaraj G, Biochim. Biophys. Acta, 1770, 1360 (2007)

[23] Patterson GH, Knobel SM, Sharif WD, Kain SR, Piston DW, Biophys. J., 73, 2782 (1997)

[24] Chalfie M, Tu Y, Euskirchen G, Ward WW, Prasher DC, Science, 263(5148), 802 (1994)