The novel extremely psychrophilic luciferase from Metridia longa: Properties of a high-purity protein produced in insect cells

Тип публикации: статья из журнала

Год издания: 2017

Идентификатор DOI: 10.1016/j.bbrc.2016.12.067

Ключевые слова: Bioluminescence, Bioluminescent reporter, Coelenterazine, Molecular adaptation, Psychrophilic enzyme

Аннотация: The bright bioluminescence of copepod Metridia longa is conditioned by a small secreted coelenterazine-dependent luciferase (MLuc). To date, three isoforms of MLuc differing in length, sequences, and some properties were cloned and successfully applied as high sensitive bioluminescent reporters. In this work, we report cloning of a novel group of genes from M. longa encoding extremely psychrophilic isoforms of MLuc (MLuc2-type). The novel isoforms share only ?54–64% of protein sequence identity with the previously cloned isoforms and, consequently, are the product of a separate group of paralogous genes. The MLuc2 isoform with consensus sequence was produced as a natively folded protein using baculovirus/insect cell expression system, purified, and characterized. The MLuc2 displays a very high bioluminescent activity and high thermostability similar to those of the previously characterized M. longa luciferase isoform MLuc7. However, in contrast to MLuc7 revealing the highest activity at 12–17 °C and 0.5 M NaCl, the bioluminescence optima of MLuc2 isoforms are at ?5 °C and 1 M NaCl. The MLuc2 adaptation to cold is also accompanied by decrease of melting temperature and affinity to substrate suggesting a more conformational flexibility of a protein structure. The luciferase isoforms with different temperature optima may provide adaptability of the M. longa bioluminescence to the changes of water temperature during diurnal vertical migrations. © 2016 Elsevier Inc.

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Издание

Журнал: Biochemical and Biophysical Research Communications

Выпуск журнала: Vol. 483, Is. 1

Номера страниц: 772-778

ISSN журнала: 0006291X

Авторы

  • Larionova Marina D. (Fed Res Ctr Krasnoyarsk Sci Ctr SB RAS, Inst Biophys SB RAS, Photobiol Lab, Krasnoyarsk, Russia; Siberian Fed Univ, Krasnoyarsk, Russia)
  • Markova Svetlana V. (Fed Res Ctr Krasnoyarsk Sci Ctr SB RAS, Inst Biophys SB RAS, Photobiol Lab, Krasnoyarsk, Russia; Siberian Fed Univ, Krasnoyarsk, Russia)
  • Vysotski Eugene S. (Fed Res Ctr Krasnoyarsk Sci Ctr SB RAS, Inst Biophys SB RAS, Photobiol Lab, Krasnoyarsk, Russia; Siberian Fed Univ, Krasnoyarsk, Russia)

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