Synthetic Control of Green Fluorescent Protein Export

@article{citeulike:5978344, abstract = {Semisynthetic green fluorescent proteins (GFPs) can be prepared by producing truncated GFPs recombinantly and assembling them with synthetic β-strands of GFP. The yield from expressing the truncated GFPs is low, and the chromophore is either partially formed or not formed. An alternative method is presented in which full-length proteins are produced recombinantly with a protease site inserted between the structural element to be removed and the rest of the protein. The native peptide can then be replaced by cutting the protease site with trypsin, denaturing in guanidine hydrochloride to disrupt the complex, separating the native peptide from the rest of the protein by size exclusion, and refolding the protein in the presence of a synthetic peptide. We show that this method allows for removal and replacement of the interior chromophore containing helix and that the GFP barrel is capable of inducing chromophore formation in a synthetic interior helix.}, author = {Kent, Kevin P. and Oltrogge, Luke M. and Boxer, Steven G.}, citeulike-article-id = {5978344}, citeulike-linkout-0 = {http://dx.doi.org/10.1021/ja906303f}, citeulike-linkout-1 = {http://pubs.acs.org/doi/abs/10.1021/ja906303f}, doi = {10.1021/ja906303f}, journal = {Journal of the American Chemical Society}, keywords = {gfp}, number = {0}, posted-at = {2009-10-21 03:38:31}, priority = {2}, title = {Synthetic Control of Green Fluorescent Protein}, url = {http://dx.doi.org/10.1021/ja906303f}, volume = {0}, year = {0000} }

TY - JOUR ID - citeulike:5978344 L3 - citeulike-article-id:5978344 N2 - Semisynthetic green fluorescent proteins (GFPs) can be prepared by producing truncated GFPs recombinantly and assembling them with synthetic β-strands of GFP. The yield from expressing the truncated GFPs is low, and the chromophore is either partially formed or not formed. An alternative method is presented in which full-length proteins are produced recombinantly with a protease site inserted between the structural element to be removed and the rest of the protein. The native peptide can then be replaced by cutting the protease site with trypsin, denaturing in guanidine hydrochloride to disrupt the complex, separating the native peptide from the rest of the protein by size exclusion, and refolding the protein in the presence of a synthetic peptide. We show that this method allows for removal and replacement of the interior chromophore containing helix and that the GFP barrel is capable of inducing chromophore formation in a synthetic interior helix. IS - 0 JF - Journal of the American Chemical Society TI - Synthetic Control of Green Fluorescent Protein VL - 0 KW - gfp AU - Kent, Kevin AU - Oltrogge, Luke AU - Boxer, Steven PY - 2000/// UR - http://dx.doi.org/10.1021/ja906303f ER -