Renaturation of Aequorea green-fluorescent protein

doi:10.1016/0006-291X(81)91599-0

Biochemical and Biophysical Research Communications

Volume 101, Issue 4, 31 August 1981, Pages 1372-1380

https://doi.org/10.1016/0006-291X(81)91599-0 Get rights and content

Abstract

Renaturation of $Aequorea$ green-fluorescent protein (A-GFP) was achieved for the first time following denaturation in guanidine-HCl or acid. Denaturation was accompanied by the concerted loss of visible fluorescence, alteration of absorption characteristics, and large negative deflection of CD signal in the far UV. Dialysis of a guanidine-denatured sample at pH 8 resulted in 64% renaturation (return to native absorption) and neutralization of an acid-denatured sample restored 90% of the native absorption. Renatured GFP is highly fluorescent and indistinguishable from native GFP with respect to the shape of excitation and emission spectra. Both native and denatured proteins exhibit resistance to trypsin hydrolysis and have identically broad pH and heat stability profiles, all of which suggest full renaturation.

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Renaturation of Aequorea green-fluorescent protein

Abstract

Methods Enzymol

FEBS Lett

Adv. Prot. Chem

J. Biol. Chem

Biochemistry

Biochemistry

J. Cell. Physiol

Renaturation of $Aequorea$ green-fluorescent protein