Violet and greenish photoprotein obelin mutants for reporter applications in dual-color assay

Тип публикации: статья из журнала

Год издания: 2008

Идентификатор DOI: 10.1007/s00216-008-2223-5

Ключевые слова: Ca(2+)-regulated photoprotein, bioluminescence, dual-color assay, Bioluminescence, Ca2+-regulated photoprotein, Dual-color assay, Bioluminescence, Ca-regulated photoprotein, Dual-color assay, Photoprotein, Calcium, calcium, follitropin, luteinizing hormone, obelin, photoprotein, article, binding site, chemistry, color, genetics, immunoassay, luminescence, methodology, point mutation, time, Binding Sites, Calcium, Color, Follicle Stimulating Hormone, Immunoassay, Luminescent Measurements, Luminescent Proteins, Luteinizing Hormone, Point Mutation, Time Factors

Аннотация: Two kinds of Ca(2+)-regulated photoprotein obelin with altered color of bioluminescence were obtained by active-center amino acid substitution. The mutant W92F-H22E emits violet light (lambda(max)=390 nm) and the mutant Y139F emits greenish light (lambda (max)=498 nm), with small spectral overlap, both display high activity and stability and thus may be used as reporters. For demonstration, the mutants were applied in dual-color simultaneous immunoassay of two gonadotropic hormones-follicle-stimulating hormone and luteinizing hormone. Bioluminescence of the reporters was simultaneously triggered by single injection of Ca(2+) solution, divided using band-pass optical filters and measured with a two-channel photometer. The sensitivity of simultaneous bioluminescence assay was close to that of a separate radioimmunoassay.

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Издание

Журнал: ANALYTICAL AND BIOANALYTICAL CHEMISTRY

Выпуск журнала: Vol. 391, Is. 8

Номера страниц: 2891-2896

ISSN журнала: 16182642

Место издания: HEIDELBERG

Издатель: SPRINGER HEIDELBERG

Авторы

  • Frank L.A. (Siberian Federal University)
  • Borisova V.V. (Institute of Biophysics,Russian Academy of Sciences,Siberian Branch)
  • Markova S.V. (Institute of Biophysics,Russian Academy of Sciences,Siberian Branch)
  • Malikova N.P. (Institute of Biophysics,Russian Academy of Sciences,Siberian Branch)
  • Stepanyuk G.A. (Institute of Biophysics,Russian Academy of Sciences,Siberian Branch)
  • Vysotski E.S. (Institute of Biophysics,Russian Academy of Sciences,Siberian Branch)

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