Engineering highly stable variants of Corynactis californica green fluorescent proteins.

Autor: Hung LW; Bioscience Division, MS M888, Los Alamos National Laboratory, Los Alamos, New Mexico, USA., Terwilliger TC; Bioscience Division, MS M888, Los Alamos National Laboratory, Los Alamos, New Mexico, USA.; New Mexico Consortium, Los Alamos, New Mexico, USA., Waldo GS; Bioscience Division, MS M888, Los Alamos National Laboratory, Los Alamos, New Mexico, USA., Nguyen HB; Bioscience Division, MS M888, Los Alamos National Laboratory, Los Alamos, New Mexico, USA.
Jazyk: angličtina
Zdroj: Protein science : a publication of the Protein Society [Protein Sci] 2024 Feb; Vol. 33 (2), pp. e4886.
DOI: 10.1002/pro.4886
Abstrakt: 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.
(© 2023 The Authors. Protein Science published by Wiley Periodicals LLC on behalf of The Protein Society.)
Databáze: MEDLINE
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