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Free phycocyanobilin (PCB) is useful in structure/function studies of phytochrome photoreceptors. Characteristically, apophytochromes can attach bilin cofactors such as PCB autocatalytically in vitro, thereby allowing chemical modification or separate isotopic labeling for vibrational spectroscopy, in particular NMR. Here we describe methods for producing, purifying, and analyzing unlabeled and universally 13C/15N-labeled PCB from cyanobacteria. We also outline methods for in vitro autoassembly and analysis of holophytochromes from apoprotein and PCB.

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Preparation of Phycocyanobilin (PCB) with and without 13C/15N Isotope Labeling for in vitro Phytochrome Autoassembly

  • 实验方案

  • 发表日期：

    2026年

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  • Jon Hughes,
  • Christina Lang

摘要

Free phycocyanobilin (PCB) is useful in structure/function studies of phytochrome photoreceptors. Characteristically, apophytochromes can attach bilin cofactors such as PCB autocatalytically in vitro, thereby allowing chemical modification or separate isotopic labeling for vibrational spectroscopy, in particular NMR. Here we describe methods for producing, purifying, and analyzing unlabeled and universally 13C/15N-labeled PCB from cyanobacteria. We also outline methods for in vitro autoassembly and analysis of holophytochromes from apoprotein and PCB.