8U21

A Highly Stable Variant of Corynactis Californica Green Fluorescent Protein, ccGFP E6

Summary for 8U21

• Entry DOI: 10.2210/pdb8u21/pdb
• Descriptor: Green Fluorescent Protein Variant E6, ccGFP E6 (2 entities in total)
• Functional Keywords: florescent protein, fluorescent protein
• Biological source: Corynactis californica
• Total number of polymer chains: 1
• Total formula weight: 24865.23

Authors

Hung, L.-W.,Terwilliger, T.C.,Waldo, G.,Nguyen, H.B. (deposition date: 2023-09-05, release date: 2024-01-31, Last modification date: 2024-10-09)

Primary citation

Hung, L.W.,Terwilliger, T.C.,Waldo, G.S.,Nguyen, H.B.
Engineering highly stable variants of Corynactis californica green fluorescent proteins.
Protein Sci., 33:e4886-e4886, 2024

PubMed Abstract: Fluorescent proteins (FPs) are versatile biomarkers that facilitate effective detection and tracking of macromolecules of interest in real time. Engineered FPs such as superfolder green fluorescent protein (sfGFP) and superfolder Cherry (sfCherry) have exceptional refolding capability capable of delivering fluorescent readout in harsh environments where most proteins lose their native functions. Our recent work on the development of a split FP from a species of strawberry anemone, Corynactis californica, delivered pairs of fragments with up to threefold faster complementation than split GFP. We present the biophysical, biochemical, and structural characteristics of five full-length variants derived from these split C. californica GFP (ccGFP). These ccGFP variants are more tolerant under chemical denaturation with up to 8 kcal/mol lower unfolding free energy than that of the sfGFP. It is likely that some of these ccGFP variants could be suitable as biomarkers under more adverse environments where sfGFP fails to survive. A structural analysis suggests explanations of the variations in stabilities among the ccGFP variants.

PubMed: 38151801
DOI: 10.1002/pro.4886

Experimental method

X-RAY DIFFRACTION (1.96 Å)