Engineering highly stable variants of Corynactis californica green fluorescent proteins

Hung, Li‐Wei; Terwilliger, Thomas C.; Waldo, Geoffrey S.; Nguyen, Hau B.

doi:10.1002/pro.4886

Engineering highly stable variants of Corynactis californica green fluorescent proteins

Journal Article · Tue Dec 26 23:00:00 EST 2023 · Protein Science

DOI:https://doi.org/10.1002/pro.4886· OSTI ID:2329254

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Los Alamos National Laboratory (LANL), Los Alamos, NM (United States). Bioscience Division
Los Alamos National Laboratory (LANL), Los Alamos, NM (United States). Bioscience Division; New Mexico Consortium, Los Alamos, NM (United States)

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.

View Accepted Manuscript (DOE)

Research Organization:: Los Alamos National Laboratory (LANL), Los Alamos, NM (United States)

Sponsoring Organization:: USDOE National Nuclear Security Administration (NNSA); USDOE Laboratory Directed Research and Development (LDRD) Program; National Institutes of Health (NIH); USDOE Office of Science (SC)

Grant/Contract Number:: 89233218CNA000001

OSTI ID:: 2329254

Report Number(s):: LA-UR--23-30129

Journal Information:: Protein Science, Journal Name: Protein Science Journal Issue: 2 Vol. 33; ISSN 0961-8368

Publisher:: WileyCopyright Statement

Country of Publication:: United States

Language:: English

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59 BASIC BIOLOGICAL SCIENCES
denaturation
green fluorescent protein
protein fragment complementation
protein detection
protein expression
protein stability
protein tagging
solubility
stable

Engineering highly stable variants of Corynactis californica green fluorescent proteins

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