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Engineering highly stable variants of Corynactis californica green fluorescent proteins.

Authors :
Hung, Li‐Wei
Terwilliger, Thomas C.
Waldo, Geoffrey S.
Nguyen, Hau B.
Source :
Protein Science: A Publication of the Protein Society; Feb2024, Vol. 33 Issue 2, p1-12, 12p
Publication Year :
2024

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. [ABSTRACT FROM AUTHOR]

Details

Language :
English
ISSN :
09618368
Volume :
33
Issue :
2
Database :
Complementary Index
Journal :
Protein Science: A Publication of the Protein Society
Publication Type :
Academic Journal
Accession number :
175071434
Full Text :
https://doi.org/10.1002/pro.4886