A protein switch sensing system for the quantification of sulfate

Krystal Teasley Hamorsky, Charles Mark Ensor, Patrizia Pasini, Sylvia Daunert

Research output: Contribution to journalArticle

7 Scopus citations

Abstract

Protein engineering has generated versatile methods and technologies that have been instrumental in advancements in the fields of sensing, therapeutics, and diagnostics. Herein, we demonstrate the employment of rational design to engineer a unique bioluminescence-based protein switch. A fusion protein switch combines two totally unrelated proteins, with distinct characteristics, in a manner such that the function of one protein is dependent on another. Herein we report a protein switch sensing system by insertion of the sulfate-binding protein (SBP) into the structure of the photoprotein aequorin (AEQ). In the presence of sulfate, SBP undergoes a conformational change bringing the two segments of AEQ together, "turning on" bioluminescence in a dose-dependent fashion, thus allowing quantitative detection of sulfate. A calibration plot was obtained by correlating the amount of bioluminescence generated with the concentration of sulfate present. The switch demonstrated selectivity and reproducibility, and a detection limit of 1.6 × 10 -4 M for sulfate. Moreover, the sensing system was validated by performing sulfate detection in clinical and environmental samples, such as, serum, urine, and tap water. The detection limits and working ranges in all three samples fall within the average normal/recommended sulfate levels in the respective matrices.

Original languageEnglish (US)
Pages (from-to)172-180
Number of pages9
JournalAnalytical Biochemistry
Volume421
Issue number1
DOIs
StatePublished - Feb 1 2012

Keywords

  • Binding protein
  • Photoprotein
  • Protein engineering
  • Protein switch
  • Sensing system

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology
  • Cell Biology

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