Aequorin mutants with increased thermostability

Xiaoge Qu, Laura Rowe, Emre Dikici, Mark Ensor, Sylvia Daunert

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Bioluminescent labels can be especially useful for in vivo and live animal studies due to the negligible bioluminescence background in cells and most animals, and the non-toxicity of bioluminescent reporter systems. Significant thermal stability of bioluminescent labels is essential, however, due to the longitudinal nature and physiological temperature conditions of many bioluminescent-based studies. To improve the thermostability of the bioluminescent protein aequorin, we employed random and rational mutagenesis strategies to create two thermostable double mutants, S32T/E156V and M36I/E146K, and a particularly thermostable quadruple mutant, S32T/E156V/Q168R/L170I. The double aequorin mutants, S32T/E156V and M36I/E146K, retained 4 and 2.75 times more of their initial bioluminescence activity than wild-type aequorin during thermostability studies at 37 °C. Moreover, the quadruple aequorin mutant, S32T/E156V/Q168R/L170I, exhibited more thermostability at a variety of temperatures than either double mutant alone, producing the most thermostable aequorin mutant identified thus far.

Original languageEnglish (US)
Pages (from-to)5639-5643
Number of pages5
JournalAnalytical and Bioanalytical Chemistry
Volume406
Issue number23
DOIs
StatePublished - Sep 1 2014
Externally publishedYes

Fingerprint

Aequorin
Bioluminescence
Labels
Animals
Luminescent Proteins
Mutagenesis
Temperature
Thermodynamic stability
Hot Temperature

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry

Cite this

Aequorin mutants with increased thermostability. / Qu, Xiaoge; Rowe, Laura; Dikici, Emre; Ensor, Mark; Daunert, Sylvia.

In: Analytical and Bioanalytical Chemistry, Vol. 406, No. 23, 01.09.2014, p. 5639-5643.

Research output: Contribution to journalArticle

Qu, Xiaoge ; Rowe, Laura ; Dikici, Emre ; Ensor, Mark ; Daunert, Sylvia. / Aequorin mutants with increased thermostability. In: Analytical and Bioanalytical Chemistry. 2014 ; Vol. 406, No. 23. pp. 5639-5643.
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